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Additional Supporting Information may be found in the online version of this article.

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HEP_25871_sm_SuppFig1.tif8636KSupporting Information Figure 1. Genotype of the LDLR gene in control and ‘JD’ pluripotent stem cells. (a) PCR of genomic DNA using primers that span deletion FH381 in the LDLR gene yields the expected 174 bp amplicon exclusively in JD iPSCs (JD1, JD3, JD4). This product is not amplified from control cells (H1 and H9 ESC lines and K3 iPSC line) due to the presence of 5kb of intervening sequence that lies between the amplification primers. (b) DNA sequence analyses confirmed the presence of the ‘JD’ paternal LDLR mutation (FH382, A-G transition) in genomic DNA from ‘JD’ fibroblasts and iPSCs, whereas only the wild type SNP was detected in control pluripotent stem cells (H1, H9 and K3).
HEP_25871_sm_SuppFig2.tif9159KSupporting Information Figure 2. FL-LDL uptake in ‘JD’ iPSC derived hepatocytes. Following differentiation and overnight incubation with 0.5μM lovastatin, control and ‘JD’ stem cell-derived hepatocytes were incubated for 30 mins with FL-LDL. Cells were fixed and the location of FL-LDL was identified by confocal microscopy. The cell location and plate orientation was marked and hepatocyte transcription factor HNF4a (red) was identified in the nuclei of the same cells by immunocytochemistry. Scale bar = 50μM.
HEP_25871_sm_SuppFig3.tif4922KSupporting Information Figure 3. qRT-PCR analyses of mRNAs encoding proteins involved in the activation of lovastatin lactone. Bar graphs showing that no significant difference in the level of mRNAs encoding carboxyesterases 1 and 2 (CES1, CES2), paraoxonases 2 and 3 (PON2, PON3), or CYP3A4 were detected between control (green bars) and ‘JD’ (red bars) hepatocytes. Error bars = ±SEM, n = 3 independent differentiations. Significance determined by 2-sided t-test, independent samples.
HEP_25871_sm_SuppFig4.tif7229KSupporting Information Figure 4. qRT-PCR analyses of mRNAs encoding sterol response element binding proteins (SREBP) (a) Bar graphs showing relative levels of SREBP1 and SREBP2 mRNAs in control (green bars) and ‘JD’ (red bars) hepatocytes that were determined by qRT-PCR. No significant difference in mRNA levels was identified between control and ‘JD’. (b) Bar graphs showing levels of SREBP1 and SREBP2 mRNA in control (green bars) and ‘JD’ (red bars) hepatocytes following lovastatin treatment. All values were normalized to the levels of HNF4a mRNA to account for subtle variances in differentiation efficiency. Graphs are presented as the change in mRNA level found in lovastatin treated cells compared to untreated cells. Error bars = ±SEM, n = 3 independent differentiations for all samples except H9 +lovastatin and JD1 +lovastatin, where n=2. Significance determined by 2-sided t-test, independent samples.
HEP_25871_sm_SuppFig5.tif1876KSupporting Information Figure 5. Rate of Fl-LDL uptake in iPSC- and ESC-derived hepatocytes. Hepatocytes generated from H1 (open triangles) or H9 (solid squares) ESCs were washed and labeled with FL-LDL for the indicated time at 37 °C at which point cells were treated with heparin and collected for flow cytometry. Error bars = SD, n = 2 biological replicates per cell line per time point.
HEP_25871_sm_SuppInfo.doc72KSupporting Information

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