Effect of siRNAs, TGF-β1 and/or PDGF-AB on HSC proliferation.

<p>(A-B) hTERT-HSCs (A) and primary HSCs (B) were transfected for 72 hours with siNrf2, siKeap1 or siCON. Cells were then detached and plated for EdU staining as described in Materials and methods section. The pictures were taken with 10X magnification by using confocal microscopy and nuclei were counted with Image J software. siCON: scrambled siRNA; siNrf2: Nrf2 knockdown; siKeap1: Keap1 knockdown. *, P ≤ 0.05; **, P ≤ 0.01; *** P ≤ 0.001 vs control or siCON. Values are expressed as rate of proliferation (mean ± SD); N = 4 independent experiment with 5 replicates each for hTERT-HSC, and N = 6 different batches with 5 replicates each. (C-D) Proliferation rate of transfected hTERT-HSC (C) and primary HSC (D) after exposure to 0.5–1 ng/mL TGF-β1 and/or 1–5 ng/mL PDGF-AB for 48 hours. EdU was added during the last 30 hours of the experiment. Values are expressed as percentage of proliferation over each control sample. *, P ≤ 0.05; **, P ≤ 0.01; *** P ≤ 0.001 vs Control (mean ± SD, N = 4 replicates for hTERT-HSC and N = 3 from different batches with 4 replicates each for primary HSC).