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Wenxi Yu, Joshua Daniel, Dhara Mehta, Krishna Rao Maddipati, Miriam L. Greenberg
MCK1 is a novel regulator of myo-inositol phosphate synthase (MIPS) that is required for inhibition of inositol synthesis by the mood stabilizer valproate
www.plosone.org
www.plosone.org
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(7)Tj
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([)Tj
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(17)Tj
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(18)Tj
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(8)Tj
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(18)Tj
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(17)Tj
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(inositol)Tj
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3.9798 0 Td
(hypothesis)Tj
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(drugs,)Tj
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(inhibition)Tj
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(mechanism)Tj
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3.4639 0 Td
(was)Tj
1.7007 0 Td
(first)Tj
1.8029 0 Td
(reported)Tj
3.6623 0 Td
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1.0261 0 Td
(cause)Tj
2.3868 0 Td
(GSK3)Tj
/F16 1 Tf
()Tj
/F11 1 Tf
3.0501 0 Td
(inhibition)Tj
4.2236 0 Td
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/F10 1 Tf
-34.8319 -1.2982 Td
[()25(\n)2()6()11()11()-4()]TJ
/F11 1 Tf
3.5716 0 Td
([)Tj
0.83 0.64 0.02 0 k
(19)Tj
0 g
(].)Tj
2.0749 0 Td
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(of)Tj
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(lithium)Tj
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1.7631 0 Td
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(model)Tj
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([)Tj
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(20)Tj
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()Tj
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(24)Tj
0 g
(].)Tj
1.7347 0 Td
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5.4936 0 Td
(VPA)Tj
2.1543 0 Td
(has)Tj
1.5477 0 Td
(also)Tj
1.7631 0 Td
(been)Tj
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1.0261 0 Td
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2.9026 0 Td
(GSK3)Tj
2.5625 0 Td
(in)Tj
1.0319 0 Td
(several)Tj
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(studies)Tj
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([)Tj
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(25)Tj
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()Tj
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(28)Tj
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(account)Tj
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(serine/threonine)Tj
6.8598 0 Td
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(29)Tj
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()Tj
0.83 0.64 0.02 0 k
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(31)Tj
0 g
(].)Tj
1.7347 0 Td
(GSK3)Tj
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()Tj
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3.1295 0 Td
(and)Tj
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(GSK3)Tj
/F16 1 Tf
()Tj
/F11 1 Tf
(,)Tj
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2.6476 0 Td
(contain)Tj
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(32)Tj
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(,)Tj
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1.7405 0 Td
(33)Tj
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(].)Tj
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(34)Tj
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Q
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BT
/F11 1 Tf
1 TL
-0.009 Tc
0 0 Td
(C)Tj
0.8787 0 Td
(labeled)Tj
2.9934 0 Td
(inositol-)Tj
-34.1119 -1.2982 Td
(3-phosphate)Tj
5.1024 0 Td
(\([)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 257.7259 659.8488 cm
BT
/F11 1 Tf
1 TL
-0.012 Tc
0 0 Td
(13)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 264.7559 655.4267 cm
BT
/F11 1 Tf
1 TL
-0.009 Tc
0 0 Td
(C)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 271.3322 653.7259 cm
BT
/F11 1 Tf
1 TL
-0.012 Tc
0 0 Td
(6)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 274.8472 655.4267 cm
BT
/F11 1 Tf
1 TL
-0.009 Tc
0 0 Td
(]-I-3-P\))Tj
3.2881 0 Td
(and)Tj
1.7121 0 Td
(inositol)Tj
3.1862 0 Td
(\([)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 363.4015 659.8488 cm
BT
/F11 1 Tf
1 TL
-0.012 Tc
0 0 Td
(13)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 370.3748 655.4267 cm
BT
/F11 1 Tf
1 TL
-0.009 Tc
0 0 Td
(C)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 377.0078 653.7259 cm
BT
/F11 1 Tf
1 TL
-0.012 Tc
0 0 Td
(6)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 380.4661 655.4267 cm
BT
/F11 1 Tf
1 TL
-0.009 Tc
0 0 Td
(]-Inositol\))Tj
4.2746 0 Td
(by)Tj
1.1678 0 Td
(LC-MS.)Tj
3.3392 0 Td
([)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 471.6283 659.8488 cm
BT
/F11 1 Tf
1 TL
-0.012 Tc
0 0 Td
(13)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 478.6582 655.4267 cm
BT
/F11 1 Tf
1 TL
-0.009 Tc
0 0 Td
(C)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 485.2346 653.7259 cm
BT
/F11 1 Tf
1 TL
-0.012 Tc
0 0 Td
(6)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 488.7496 655.4267 cm
BT
/F11 1 Tf
1 TL
-0.009 Tc
0 0 Td
(]-I-3-P)Tj
2.948 0 Td
(and)Tj
1.7177 0 Td
([)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 538.7527 659.8488 cm
BT
/F11 1 Tf
1 TL
-0.012 Tc
0 0 Td
(13)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 545.7259 655.4267 cm
BT
/F11 1 Tf
1 TL
-0.009 Tc
0 0 Td
(C)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 552.359 653.7259 cm
BT
/F11 1 Tf
1 TL
-0.012 Tc
0 0 Td
(6)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 555.8173 655.4267 cm
BT
/F11 1 Tf
1 TL
-0.009 Tc
0 0 Td
(]-)Tj
-35.5802 -1.3039 Td
(Inositol)Tj
3.2542 0 Td
(were)Tj
2.1032 0 Td
(quantified)Tj
4.2577 0 Td
(by)Tj
1.1735 0 Td
(LC-MS)Tj
3.1124 0 Td
(as)Tj
1.0035 0 Td
(follows:)Tj
3.2485 0 Td
(Samples)Tj
3.4582 0 Td
(were)Tj
2.1033 0 Td
(subjected)Tj
3.9231 0 Td
(to)Tj
1.0148 0 Td
(HPLC)Tj
2.7326 0 Td
(on)Tj
1.2642 0 Td
(a)Tj
0.6463 0 Td
(Diamond)Tj
-33.2955 -1.2982 Td
(Hydride)Tj
3.5206 0 Td
(column)Tj
3.2655 0 Td
(\(Cogent)Tj
3.4299 0 Td
(HPLC)Tj
2.7269 0 Td
(Columns,)Tj
4.0819 0 Td
(Microsolv)Tj
4.2066 0 Td
(Technology)Tj
4.8812 0 Td
(Corporation,)Tj
5.3801 0 Td
(2)Tj
0.686 0 Td
(mm)Tj
1.8538 0 Td
(x)Tj
0.6804 0 Td
(150)Tj
-34.7129 -1.2983 Td
(mm,)Tj
2.0693 0 Td
(4)Tj
/F18 1 Tf
[()]TJ
/F11 1 Tf
(,)Tj
1.4116 0 Td
(100)Tj
/F18 1 Tf
1.6328 0 Td
[()]TJ
/F11 1 Tf
(\))Tj
1.2359 0 Td
(using)Tj
2.364 0 Td
(gradient)Tj
3.515 0 Td
(elution.)Tj
3.2201 0 Td
(The)Tj
1.7688 0 Td
(gradient)Tj
3.515 0 Td
(between)Tj
3.4922 0 Td
(mobile)Tj
2.9651 0 Td
(phases)Tj
2.8119 0 Td
(A:)Tj
1.1168 0 Td
(acetonitrile-)Tj
-31.1185 -1.3039 Td
(aqueous)Tj
3.4696 0 Td
(ammonium)Tj
4.9266 0 Td
(formate)Tj
3.3222 0 Td
(\(9:1)Tj
1.7177 0 Td
(v/v\))Tj
1.7802 0 Td
(and)Tj
1.7178 0 Td
(B:)Tj
1.0148 0 Td
(acetonitrile-0.1%)Tj
6.9278 0 Td
(aqueous)Tj
3.4696 0 Td
(formic)Tj
2.8686 0 Td
(acid)Tj
1.8482 0 Td
(\(1:9)Tj
1.7178 0 Td
(v/v\))Tj
-34.7809 -1.2983 Td
(was:)Tj
1.9049 0 Td
(0)Tj
0.686 0 Td
(min)Tj
2.3527 0 Td
(95%)Tj
1.9162 0 Td
(A,)Tj
1.1168 0 Td
(2)Tj
0.6917 0 Td
(min)Tj
2.3527 0 Td
(80%)Tj
1.9162 0 Td
(A,)Tj
1.1169 0 Td
(4)Tj
0.6916 0 Td
(min)Tj
2.3528 0 Td
(30%A,)Tj
2.8119 0 Td
(7)Tj
0.6917 0 Td
(min)Tj
2.3527 0 Td
(30%A.)Tj
2.812 0 Td
(At)Tj
1.1905 0 Td
(the)Tj
1.457 0 Td
(end)Tj
1.7065 0 Td
(of)Tj
1.0034 0 Td
(each)Tj
2.0069 0 Td
(chro-)Tj
-33.1311 -1.2982 Td
(matographic)Tj
5.2271 0 Td
(run,)Tj
1.8765 0 Td
(the)Tj
1.4513 0 Td
(mobile)Tj
2.9594 0 Td
(phase)Tj
2.4604 0 Td
(composition)Tj
5.1874 0 Td
(was)Tj
1.6838 0 Td
(changed)Tj
3.5376 0 Td
(to)Tj
1.0148 0 Td
(initial)Tj
2.5114 0 Td
(conditions)Tj
4.4277 0 Td
(and)Tj
1.7122 0 Td
(equili-)Tj
-34.0496 -1.304 Td
(brated)Tj
2.7496 0 Td
(for)Tj
1.372 0 Td
(5)Tj
0.6916 0 Td
(min)Tj
1.8368 0 Td
(before)Tj
2.7099 0 Td
(the)Tj
1.457 0 Td
(start)Tj
1.9559 0 Td
(of)Tj
1.0092 0 Td
(the)Tj
1.4513 0 Td
(next)Tj
1.9389 0 Td
(sample)Tj
2.9877 0 Td
(analysis.)Tj
3.5092 0 Td
(The)Tj
1.7689 0 Td
(flow)Tj
1.9218 0 Td
(rate)Tj
1.7292 0 Td
(was)Tj
1.6837 0 Td
(0.2)Tj
1.3777 0 Td
(ml/min)Tj
3.2258 0 Td
(and)Tj
-35.3762 -1.2982 Td
(the)Tj
1.4513 0 Td
(column)Tj
3.2655 0 Td
(was)Tj
1.6838 0 Td
(maintained)Tj
4.7395 0 Td
(at)Tj
0.9411 0 Td
(35C.)Tj
2.4264 0 Td
(The)Tj
1.7688 0 Td
(eluent)Tj
2.6589 0 Td
(was)Tj
1.6838 0 Td
(directly)Tj
3.1918 0 Td
(introduced)Tj
4.5921 0 Td
(to)Tj
1.0148 0 Td
(the)Tj
1.4513 0 Td
(mass)Tj
2.177 0 Td
(analyzer)Tj
-33.0461 -1.2983 Td
(\(QTRAP5500,)Tj
5.8563 0 Td
(Sciex\))Tj
2.5852 0 Td
(via)Tj
1.3606 0 Td
(a)Tj
0.6463 0 Td
(TurboIon)Tj
4.0989 0 Td
(Electrospray)Tj
5.1364 0 Td
(ionization)Tj
4.2292 0 Td
(source.)Tj
3.0217 0 Td
(The)Tj
1.7688 0 Td
(following)Tj
3.9345 0 Td
(parameters)Tj
-32.6379 -1.3039 Td
(were)Tj
2.1033 0 Td
(used)Tj
2.0353 0 Td
(for)Tj
1.3719 0 Td
(the)Tj
1.457 0 Td
(mass)Tj
2.177 0 Td
(analyzer:)Tj
3.7247 0 Td
(Ion)Tj
1.5987 0 Td
(spray)Tj
2.3358 0 Td
(voltage:)Tj
3.2315 0 Td
(-4500)Tj
2.4491 0 Td
(V,)Tj
1.1282 0 Td
(Curtain)Tj
3.2938 0 Td
(gas:)Tj
1.6894 0 Td
(35,)Tj
1.3777 0 Td
(Source)Tj
2.9196 0 Td
(tempera-)Tj
-32.893 -1.2983 Td
(ture:)Tj
2.0409 0 Td
(600C,)Tj
2.897 0 Td
(GS1)Tj
1.8709 0 Td
(&)Tj
0.9184 0 Td
(GS2:)Tj
2.092 0 Td
(45,)Tj
1.3833 0 Td
(and)Tj
1.7177 0 Td
(EP)Tj
1.338 0 Td
(&)Tj
0.9184 0 Td
(CXP:)Tj
2.3017 0 Td
(10.)Tj
1.3776 0 Td
([)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 391.9748 516.8692 cm
BT
/F11 1 Tf
1 TL
-0.012 Tc
0 0 Td
(13)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 398.948 512.3905 cm
BT
/F11 1 Tf
1 TL
-0.009 Tc
0 0 Td
(C)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 405.5244 510.7464 cm
BT
/F11 1 Tf
1 TL
-0.012 Tc
0 0 Td
(6)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 409.0393 512.3905 cm
BT
/F11 1 Tf
1 TL
-0.009 Tc
0 0 Td
(]-I-3-P)Tj
2.9536 0 Td
(and)Tj
1.7121 0 Td
([)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 459.0425 516.8692 cm
BT
/F11 1 Tf
1 TL
-0.012 Tc
0 0 Td
(13)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 466.0724 512.3905 cm
BT
/F11 1 Tf
1 TL
-0.009 Tc
0 0 Td
(C)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 472.6488 510.7464 cm
BT
/F11 1 Tf
1 TL
-0.012 Tc
0 0 Td
(6)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 476.1637 512.3905 cm
BT
/F11 1 Tf
1 TL
-0.009 Tc
0 0 Td
(]-inositol)Tj
3.8607 0 Td
(were)Tj
2.1033 0 Td
(moni-)Tj
-33.5789 -1.2982 Td
(tored)Tj
2.3187 0 Td
(by)Tj
1.1736 0 Td
(Multiple)Tj
3.5999 0 Td
(Reaction)Tj
3.7021 0 Td
(Monitoring)Tj
4.8415 0 Td
(with)Tj
1.9786 0 Td
(the)Tj
1.457 0 Td
(following)Tj
3.9344 0 Td
(transitions,)Tj
4.6431 0 Td
(declustering)Tj
5.0456 0 Td
(potentials)Tj
-32.6945 -1.2983 Td
(\(DP\),)Tj
2.3924 0 Td
(and)Tj
1.7178 0 Td
(collision)Tj
3.549 0 Td
(energies)Tj
3.4639 0 Td
(\(CE\).)Tj
2.33 0 Td
([)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 337.8897 490.8472 cm
BT
/F11 1 Tf
1 TL
-0.012 Tc
0 0 Td
(13)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 344.9196 486.4251 cm
BT
/F11 1 Tf
1 TL
-0.009 Tc
0 0 Td
(C)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 351.496 484.7244 cm
BT
/F11 1 Tf
1 TL
-0.012 Tc
0 0 Td
(6)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 355.011 486.4251 cm
BT
/F11 1 Tf
1 TL
-0.009 Tc
0 0 Td
(]-I-3-P)Tj
/F10 1 Tf
2.9536 0 Td
[( !")]TJ
/F11 1 Tf
1.7008 0 Td
(265.1)Tj
2.3244 0 Td
(to)Tj
1.0148 0 Td
(79)Tj
1.1622 0 Td
(at)Tj
0.9411 0 Td
(DP:)Tj
1.7291 0 Td
(-110)Tj
1.9729 0 Td
(V)Tj
0.9128 0 Td
(and)Tj
1.7121 0 Td
(CE:)Tj
1.6667 0 Td
(-25)Tj
1.5024 0 Td
(V;)Tj
-35.0926 -1.3039 Td
([)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 203.3574 477.8645 cm
BT
/F11 1 Tf
1 TL
-0.012 Tc
0 0 Td
(13)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 210.3874 473.3858 cm
BT
/F11 1 Tf
1 TL
-0.009 Tc
0 0 Td
(C)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 216.9637 471.7417 cm
BT
/F11 1 Tf
1 TL
-0.012 Tc
0 0 Td
(6)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 220.4787 473.3858 cm
BT
/F11 1 Tf
1 TL
-0.009 Tc
0 0 Td
(]-inositol)Tj
/F10 1 Tf
3.8607 0 Td
[( !")]TJ
/F11 1 Tf
1.7065 0 Td
(185.1)Tj
2.3187 0 Td
(to)Tj
1.0148 0 Td
(89)Tj
1.1622 0 Td
(at)Tj
0.9411 0 Td
(DP:)Tj
1.7291 0 Td
(-96)Tj
1.508 0 Td
(V)Tj
0.9071 0 Td
(and)Tj
1.7178 0 Td
(CE:)Tj
1.6611 0 Td
(-21)Tj
1.508 0 Td
(V.)Tj
1.1282 0 Td
(Under)Tj
2.8006 0 Td
(these)Tj
2.228 0 Td
(conditions,)Tj
4.6488 0 Td
(the)Tj
1.4513 0 Td
(reten-)Tj
-34.3386 -1.2982 Td
(tion)Tj
1.8199 0 Td
(times)Tj
2.364 0 Td
(for)Tj
1.3719 0 Td
([)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 258.9165 464.8251 cm
BT
/F11 1 Tf
1 TL
-0.012 Tc
0 0 Td
(13)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 265.9464 460.4031 cm
BT
/F11 1 Tf
1 TL
-0.009 Tc
0 0 Td
(C)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 272.5228 458.759 cm
BT
/F11 1 Tf
1 TL
-0.012 Tc
0 0 Td
(6)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 276.0377 460.4031 cm
BT
/F11 1 Tf
1 TL
-0.009 Tc
0 0 Td
(]-I-3-P)Tj
2.948 0 Td
(and)Tj
1.7177 0 Td
([)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 326.0409 464.8251 cm
BT
/F11 1 Tf
1 TL
-0.012 Tc
0 0 Td
(13)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 333.0708 460.4031 cm
BT
/F11 1 Tf
1 TL
-0.009 Tc
0 0 Td
(C)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 339.6472 458.759 cm
BT
/F11 1 Tf
1 TL
-0.012 Tc
0 0 Td
(6)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 343.1622 460.4031 cm
BT
/F11 1 Tf
1 TL
-0.009 Tc
0 0 Td
(]-inositol)Tj
3.8607 0 Td
(were)Tj
2.1033 0 Td
(4.7)Tj
1.3776 0 Td
(and)Tj
1.7178 0 Td
(3.6)Tj
1.3833 0 Td
(min,)Tj
2.058 0 Td
(respectively.)Tj
5.0456 0 Td
(The)Tj
1.7688 0 Td
(detection)Tj
-33.6299 -1.2982 Td
(limits)Tj
2.4491 0 Td
(were)Tj
2.1033 0 Td
(0.1)Tj
1.3776 0 Td
(ng)Tj
1.2246 0 Td
(and)Tj
1.7178 0 Td
(the)Tj
1.4513 0 Td
(quantitation)Tj
5.0966 0 Td
(limits)Tj
2.4491 0 Td
(were)Tj
2.1033 0 Td
(0.4)Tj
1.3833 0 Td
(ng)Tj
1.2245 0 Td
(on)Tj
1.2643 0 Td
(the)Tj
1.4513 0 Td
(column)Tj
3.2655 0 Td
(for)Tj
1.3719 0 Td
(each)Tj
2.007 0 Td
(analyte.)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 418.2803 cm
BT
/F0 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(MIPS)Tj
2.6504 0 Td
(activity)Tj
3.2457 0 Td
(assay)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 401.2724 cm
BT
/F11 1 Tf
1 TL
-0.012 Tc
0 0 Td
(Cells)Tj
2.1203 0 Td
(expressing)Tj
4.3426 0 Td
(His-Xpress)Tj
4.5751 0 Td
(tagged)Tj
2.7836 0 Td
(MIPS)Tj
2.4548 0 Td
(were)Tj
2.0919 0 Td
(grown)Tj
2.7553 0 Td
(to)Tj
1.0091 0 Td
(the)Tj
1.4457 0 Td
(early)Tj
2.1089 0 Td
(stationary)Tj
4.1159 0 Td
(phase)Tj
2.4435 0 Td
(and)Tj
1.7064 0 Td
(lysed)Tj
2.194 0 Td
(in)Tj
-36.1471 -1.2982 Td
(buffer)Tj
2.5738 0 Td
(\(50)Tj
1.491 0 Td
(mM)Tj
1.9162 0 Td
(Tris-HCl)Tj
3.7927 0 Td
(pH)Tj
1.4911 0 Td
(7.5,)Tj
1.5874 0 Td
(0.6)Tj
1.3719 0 Td
(M)Tj
1.1055 0 Td
(sorbitol,)Tj
3.4413 0 Td
(0.3)Tj
1.3719 0 Td
(M)Tj
1.1055 0 Td
(NaCl,)Tj
2.4832 0 Td
(1X)Tj
1.3322 0 Td
(protease)Tj
3.4923 0 Td
(inhibitor)Tj
3.7077 0 Td
(and)Tj
1.7064 0 Td
(1X)Tj
1.3323 0 Td
(phos-)Tj
-35.3024 -1.3039 Td
(phatase)Tj
3.1577 0 Td
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