Figure 1.
Cell extracts were prepared from hMFB, LX-2, HepG2, Hep3B, HUVEC and Wi-38 VA-13 subline 2RA and analysed in Western blot for expression of (i) extracellular matrix proteins (fibronectin, collagen IV, collagen I), (ii) receptors of TGF-β (endoglin, TβRII, TβRI), (iii) HSC/MFB markers (vimentin, desmin, α-SMA, CTGF, Id2, p21), and (iv) SV40 large T-antigen and β-actin. We used two parenchymatic (HepG2, Hep3B) and other cell entities (hMFB, HUVEC, and WI-38) as controls because they are well characterized for the expression of tested genes and served as positive or negative controls in this analysis.
Figure 2.
Representative numbers and appearance of chromosomes that was found in one LX-2 cell as obtained after Giemsa stain in lightmicroscopic analysis. The karyotype was determined to 74, XYY, +del(1)(q4?1), der(2;10)(p10;p10), +del(3)(q25)x2, +der(4)add(4)(q31), der(5)?t(5;9)(q35;q34)?del(5)(p15), der(5)add(5)(p1?3), +der(5)add(5)(p1?3)x2, +6,del(7)(p1?3), +del(7)(p1?3), +der(9)(11q23->11q21::14q32->14q11.2::9p13->9qter)x3, +i(10)(q10), −11, del(11)(p11.2), +del(12)(pter->q12::q15->q22::q24.2->qter), −13, −14, +15, +der(16)t(16;17)(q11.1;q11.2), +der(18)(18pter->18q11.2::1q25->1q32::7q22->7qter), +20, +20, +21, +21, der(22)?t(22;?)(q11.2;?), +11mar, respectively. Please note that a characteristic derivative of chromosome 18, i.e. +der(18)(18pter->18q11.2::1q25->1q32::7q22->7qter), is marked with an arrow in this figure. A characteristic derivative of chromosome 18 that contains part of chromosomes 1 and 7 was seen in three karyograms and is marked by a black arrow.
Figure 3.
Spectral karyogram (SKY) of the hepatic stellate cell line LX-2.
Shown is a representative image of LX-2 cells hybridized with a 24-color SKY probe (upper left panel). The chromosomes were counterstained with DAPI (upper right panel). The spectral images of hybridized metaphases were acquired using a cytogenetic workstation and grouped with the SkyView software (lower panel). Based on this analysis, the karyotype of this LX-2 metaphase is determined to: 73<3n+>, XXXYY, ish +der(1)t(1;18)(p11;q21)t(1;7)(q42;p12), der(2)t(2;10)(p11;q11), del(2)(q11), +der(3)t(3;6)(qter;q11), der(3)t(3;5)(p11;q23)t(3;5)(q24;q23)del(5)(q14), −5, +del(6)(p11), der(6)t(6;14)(p11;q11), der(6)t(6;11)(q11;q12), der(7)t(7;18)(p11;p11), del(7)(q21), +der(8)t(8;5)(q11;q11)t(5;14)(q31;q23), der(8)t(4;8)(q32;p12), +der(9)t(3;9)(p22;qter)del(9)(p11), der(9)t(9;14)(p12;q11)t(14;11)(q24;q14)del(11)(q12), +der(10)del(10)(p12)del(10)(q23), der(10)t(5;10)(p12;p11), del(11)(p14), der(11)del(11)(p14)del(11)(q23)x2, der(12)t(12;22)(q13;q12), −13, −14x2,+i(15)(p10), +der(15)t(15;20)(q15;q11), der(16)t(16;17)(p11;q11), del(17)(p11), −19, −20, −21, der(21)t(21;13)(p11;q11), del(X)(q24), del(X)(q25), del(X)(q21). Please note that the derivative that contains parts of chromosomes 1, 18 and 7 (cf. Figure 1) were also seen in this analysis.
Figure 4.
Median appearance of chromosome regions per cell based upon SKY analysis of eight metaphases from human LX-2 cell line.
The average copy numbers per cell of each human chromosome region are shown in correspondence to their chromosome ideogram based on summary SKY analysis.
Figure 5.
Complete SKYGRAM of LX-2 cells based on eight cells.
Characters above rearranged chromosome ideograms are for numbers of metaphases with this aberration. Rearrangements recognized only once in SKY analysis are ignored. Based on this analysis, the complete SKYGRAM of LX-2 was determined to 64∼83<n3+/−>, XXY, −1, del(1)(q11) [3], del(2)(q11) [3], +del(3)(p11) [2], del(6)(p11) [2], del(7)(p11) [4], del(7)(q11) [2], del(7)(p15) [2], +der(7)t(7;18)(p11;p11) [3], der(8)t(5;8)(q11;q11)t(5;14)(q31;q23) [4], der(9)t(9;14)(p12;q11)t(11;14)(q14;q24)del(11)(q12) [7], del(10)(q22) [4],i(10)(q10) [2], der(11)del(11)(p11)del(11)(q22)x2 [2], del(12)(q11) [2], +del(13)(q21) [2],-14,-14, +del(17)(p11) [2],+20[cp8], respectively. More detailed SKYGRAM data for LX-2 cells were deposited in the SKY/M-FISH & CGH database located at the National Center for Biotechnology Information that can be found at: http://www.ncbi.nlm.nih.gov/sky/skyquery.cgi.
Figure 6.
Chromosomal imbalances detected in the LX-2 cell line using a SNP 6.0 Array.
Gains and losses are shown by red and blue triangles, respectively.