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Lika Osugui, Jolanda J. de Roo, Vivian Cristina de Oliveira, Ana Clara Pires Sodré, Frank J. T. Staal, Ana Flavia Popi
B-1 cells and B-1 cell precursors prompt different responses to Wnt signaling
www.plosone.org
www.plosone.org
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2.1146 0 Td
(and)Tj
-33.2898 -1.2982 Td
(C57BL/6)Tj
3.685 0 Td
(wild)Tj
1.9049 0 Td
(type)Tj
/F12 1 Tf
1.8765 0 Td
[()]TJ
/F7 1 Tf
(WT\))Tj
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(mice)Tj
2.1146 0 Td
(were)Tj
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(used.)Tj
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(peritoneal)Tj
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(cells)Tj
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(harvested)Tj
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1.1622 0 Td
(washing)Tj
3.4412 0 Td
(out)Tj
1.5194 0 Td
(the)Tj
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(peritoneal)Tj
4.1556 0 Td
(cavity)Tj
2.4945 0 Td
(using)Tj
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(RPMI1640)Tj
4.456 0 Td
(medium.)Tj
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1.7518 0 Td
(bone)Tj
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(marrow)Tj
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(cells)Tj
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(isolated)Tj
3.2031 0 Td
(from)Tj
2.1656 0 Td
(both)Tj
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(femurs)Tj
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(each)Tj
1.9899 0 Td
(animal,)Tj
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(and)Tj
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(clear)Tj
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(a)Tj
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(mortar)Tj
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1.7008 0 Td
(pestle.)Tj
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(The)Tj
1.7518 0 Td
(crushed)Tj
-32.7852 -1.2983 Td
(bones)Tj
2.5172 0 Td
(were)Tj
2.0806 0 Td
(rinsed)Tj
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(RPMI1640)Tj
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(filter)Tj
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(40)Tj
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1.1509 0 Td
[()]TJ
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(m)Tj
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(cell)Tj
1.5194 0 Td
(strainer.)Tj
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(that,)Tj
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(cells)Tj
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(anti-CD16)Tj
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(CD32)Tj
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(mAb)Tj
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1.0035 0 Td
(block)Tj
2.3527 0 Td
(Fc)Tj
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(\r)Tj
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(RIII/II)Tj
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(30)Tj
1.1509 0 Td
(min)Tj
1.8255 0 Td
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1.9559 0 Td
(the)Tj
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(monoclonal)Tj
4.9096 0 Td
(antibodies)Tj
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2.9594 0 Td
(the)Tj
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3.8891 0 Td
(molecules:)Tj
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(CD19,)Tj
2.7496 0 Td
(CD23,)Tj
2.7439 0 Td
(CD11b,)Tj
3.2429 0 Td
(CD5)Tj
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(characterize)Tj
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(B-1a)Tj
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(\(CD19)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 539.8866 562.2236 cm
BT
/F7 1 Tf
1 TL
-0.0186 Tc
0 0 Td
(+)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 544.1385 557.8015 cm
BT
/F7 1 Tf
1 TL
-0.014 Tc
0 0 Td
(CD23)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 567.2692 562.2236 cm
BT
/F7 1 Tf
1 TL
-0.0186 Tc
0 0 Td
(-)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 569.7637 557.8015 cm
BT
/F7 1 Tf
1 TL
-0.014 Tc
0 0 Td
(-)Tj
-36.9748 -1.2982 Td
(CD11b)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 228.0755 549.2409 cm
BT
/F7 1 Tf
1 TL
-0.0186 Tc
0 0 Td
(+/-)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 237.2031 544.8188 cm
BT
/F7 1 Tf
1 TL
-0.014 Tc
0 0 Td
(CD5)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 255.6283 549.2409 cm
BT
/F7 1 Tf
1 TL
-0.0186 Tc
0 0 Td
(+)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 259.8803 544.8188 cm
BT
/F7 1 Tf
1 TL
-0.014 Tc
0 0 Td
(\))Tj
0.5499 0 Td
(and)Tj
1.7007 0 Td
(B-1b)Tj
2.092 0 Td
(\(CD19)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 329.7259 549.2409 cm
BT
/F7 1 Tf
1 TL
-0.0186 Tc
0 0 Td
(+)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 333.9779 544.8188 cm
BT
/F7 1 Tf
1 TL
-0.014 Tc
0 0 Td
(CD23)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 357.1086 549.2409 cm
BT
/F7 1 Tf
1 TL
-0.0186 Tc
0 0 Td
(-)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 359.6031 544.8188 cm
BT
/F7 1 Tf
1 TL
-0.014 Tc
0 0 Td
(CD11b)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 387.6661 549.2409 cm
BT
/F7 1 Tf
1 TL
-0.0186 Tc
0 0 Td
(+/-)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 396.7937 544.8188 cm
BT
/F7 1 Tf
1 TL
-0.014 Tc
0 0 Td
(CD5)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 415.2755 549.2409 cm
BT
/F7 1 Tf
1 TL
-0.0186 Tc
0 0 Td
(-)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 417.77 544.8188 cm
BT
/F7 1 Tf
1 TL
-0.014 Tc
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(\))Tj
0.5499 0 Td
(cells)Tj
1.8708 0 Td
(subsets)Tj
2.9877 0 Td
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(peritoneal)Tj
4.1612 0 Td
(cavity)Tj
2.4888 0 Td
(and)Tj
-35.9997 -1.3039 Td
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1.9559 0 Td
(CD3e,)Tj
2.6929 0 Td
(CD4,)Tj
2.2847 0 Td
(CD11b,)Tj
3.2429 0 Td
(Gr-1,)Tj
2.3073 0 Td
(IgM,)Tj
2.1033 0 Td
(NK-1.1,)Tj
3.3109 0 Td
(Ter119,)Tj
3.2088 0 Td
(CD45R/B220,)Tj
5.6465 0 Td
(CD19)Tj
2.5284 0 Td
(and)Tj
1.7008 0 Td
(CD93)Tj
2.5285 0 Td
(\(Early)Tj
2.5795 0 Td
(B)Tj
-36.0904 -1.2982 Td
(AA4.1\))Tj
3.0331 0 Td
(to)Tj
0.9978 0 Td
(characterize)Tj
4.8982 0 Td
(B-1)Tj
1.5874 0 Td
(progenitors)Tj
4.7395 0 Td
(\(B-1P)Tj
2.4605 0 Td
(-)Tj
0.5499 0 Td
(Lin)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 395.9433 523.2188 cm
BT
/F7 1 Tf
1 TL
-0.0186 Tc
0 0 Td
(-)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 398.4377 518.7968 cm
BT
/F7 1 Tf
1 TL
-0.014 Tc
0 0 Td
(CD19)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 421.5685 523.2188 cm
BT
/F7 1 Tf
1 TL
-0.0186 Tc
0 0 Td
(+)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 425.8204 518.7968 cm
BT
/F7 1 Tf
1 TL
-0.014 Tc
0 0 Td
(EarlyB)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 451.7858 523.2188 cm
BT
/F7 1 Tf
1 TL
-0.0186 Tc
0 0 Td
(+)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 455.9811 518.7968 cm
BT
/F7 1 Tf
1 TL
-0.014 Tc
0 0 Td
(B220)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 475.6535 523.2188 cm
BT
/F7 1 Tf
1 TL
-0.0186 Tc
0 0 Td
(lo/neg)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 493.9086 518.7968 cm
BT
/F7 1 Tf
1 TL
-0.014 Tc
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(\))Tj
0.5385 0 Td
(from)Tj
2.16 0 Td
(bone)Tj
2.1543 0 Td
(marrow.)Tj
-34.2422 -1.2982 Td
(B-1)Tj
1.5931 0 Td
(cells)Tj
1.8708 0 Td
(and)Tj
1.7008 0 Td
(B-1)Tj
1.5988 0 Td
(cell)Tj
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(precursors)Tj
4.354 0 Td
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2.0806 0 Td
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1.7291 0 Td
(stained)Tj
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(with)Tj
1.9559 0 Td
(anti-Flt3)Tj
3.5773 0 Td
(and)Tj
1.7008 0 Td
(IL7-R)Tj
2.4945 0 Td
(antibodies)Tj
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(deter-)Tj
-34.452 -1.3039 Td
(mine)Tj
2.2394 0 Td
(the)Tj
1.44 0 Td
(expression)Tj
4.3653 0 Td
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(these)Tj
2.2054 0 Td
(receptors.)Tj
4.0421 0 Td
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(acquired)Tj
3.617 0 Td
(using)Tj
2.3414 0 Td
(the)Tj
1.44 0 Td
(BD)Tj
1.5193 0 Td
(FACSCanto)Tj
/F14 1 Tf
[()]TJ
/F7 1 Tf
(II)Tj
6.032 0 Td
(flow)Tj
-34.4293 -1.2983 Td
(cytometer)Tj
4.1499 0 Td
(and)Tj
1.7008 0 Td
(data)Tj
1.8709 0 Td
(were)Tj
2.0862 0 Td
(analyzed)Tj
3.6227 0 Td
(with)Tj
1.9559 0 Td
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2.9593 0 Td
(software)Tj
3.5093 0 Td
(\()Tj
0.83 0.64 0.02 0 k
(S1)Tj
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1.4797 0 Td
(and)Tj
0.83 0.64 0.02 0 k
1.7008 0 Td
(S2)Tj
0 g
1.1508 0 Td
(Figs\).)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 451.2188 cm
BT
/F13 1 Tf
1 TL
-0.0033 Tc
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(of)Tj
1.0677 0 Td
(B-1)Tj
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(and)Tj
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(B-1P)Tj
2.2914 0 Td
(cells)Tj
ET
Q
q
1 j
1 J
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10 0 0 10 200.0125 434.211 cm
BT
/F7 1 Tf
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(B-1)Tj
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(cells)Tj
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(FACSAria)Tj
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(strategies)Tj
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(B-1)Tj
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(selection.)Tj
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(the)Tj
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(selection)Tj
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(CD3)Tj
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(the)Tj
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(lymphocyte)Tj
4.8926 0 Td
(gate,)Tj
2.0693 0 Td
(a)Tj
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(double)Tj
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(negative)Tj
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ET
Q
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1 j
1 J
0 w
7.4999 0 0 6.6659 240.548 386.6456 cm
BT
/F7 1 Tf
1 TL
-0.0053 Tc
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(-)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 243.1559 382.2236 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(CD23)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 266.6834 386.6456 cm
BT
/F7 1 Tf
1 TL
-0.0053 Tc
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(-)Tj
ET
Q
q
1 j
1 J
0 w
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BT
/F7 1 Tf
1 TL
-0.004 Tc
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(\))Tj
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(an)Tj
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(aliquot)Tj
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(sorted)Tj
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(cells)Tj
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(fully)Tj
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(confirm)Tj
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(the)Tj
1.4684 0 Td
(B-1)Tj
1.627 0 Td
(cell)Tj
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(purity.)Tj
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(the)Tj
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(positive)Tj
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(selection,)Tj
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(CD19)Tj
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(and)Tj
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(CD23)Tj
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(antibodies)Tj
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(were)Tj
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(used,)Tj
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(and)Tj
1.7291 0 Td
(the)Tj
-35.2627 -1.2982 Td
(CD19)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 223.4834 360.6236 cm
BT
/F7 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(+)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 227.8488 356.2015 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(CD23)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 251.3763 360.6236 cm
BT
/F7 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(-)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 256.1952 356.2015 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
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(population)Tj
4.5864 0 Td
(from)Tj
2.2053 0 Td
(the)Tj
1.474 0 Td
(lymphocyte)Tj
4.8926 0 Td
(gate)Tj
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(was)Tj
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(sorted.)Tj
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(all)Tj
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(experiments)Tj
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(the)Tj
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(B-1)Tj
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(purity)Tj
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1.7008 0 Td
(around)Tj
3.135 0 Td
(95%)Tj
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(after)Tj
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(cell)Tj
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(sorting.)Tj
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-11.8147 -1.3039 Td
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2.2337 0 Td
(marrow)Tj
3.3334 0 Td
(was)Tj
1.6668 0 Td
(processed)Tj
4.0252 0 Td
(and)Tj
1.6951 0 Td
(stained)Tj
2.9933 0 Td
(as)Tj
0.9922 0 Td
(described)Tj
3.9458 0 Td
(above)Tj
2.4831 0 Td
(\(item)Tj
2.3131 0 Td
(2\))Tj
1.0091 0 Td
(and)Tj
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(B-1P)Tj
2.1373 0 Td
(cells)Tj
1.8595 0 Td
(were)Tj
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(enriched)Tj
3.634 0 Td
(by)Tj
1.1622 0 Td
(cell)Tj
1.5079 0 Td
(sorting,)Tj
3.1691 0 Td
(using)Tj
2.3301 0 Td
(the)Tj
1.4343 0 Td
(following)Tj
3.8721 0 Td
(strategy:)Tj
3.4413 0 Td
(from)Tj
2.16 0 Td
(lineage)Tj
2.9423 0 Td
(negative)Tj
3.4412 0 Td
(population)Tj
4.4674 0 Td
(\(CD3e,)Tj
-33.5619 -1.2983 Td
(CD4,)Tj
2.2734 0 Td
(CD11b,)Tj
3.2315 0 Td
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