Fig. 5.

Ablation of BSCL2 in HepG2 cells does not alter lipid accumulation. (A) Schematic representation of CRISPR/Cas9 gRNA editing sites in exon 2 of BSCL2. (B) PCR identification of BSCL2 exon 2 deletion by CRISPR/Cas9 in a mixed population of HepG2 cells. KO, knockout; NTC, no template control; WT, wild type. (C) PCR identification of single-cell HepG2 colonies containing BSCL2 exon 2 deletion isolated by serial dilution. (D,E) PCR analysis of clones selected for expansion (D) and DNA sequencing (E) of PCR products confirming BSCL2 exon 2 deletion in HepG2 cells. (F) Western blot analysis confirming the ablation of BSCL2 protein in HepG2 single cell clones with BSCL2 exon 2 deletion. (G) Representative images of Oil Red O-stained HepG2 cell clones (scale bars: 50 µm) and quantification of the eluted stain from biological replicates (n=8); data presented are from two independent experiments. Data are presented as the mean±s.e.m., *P<0.05 vs Ctrl #7.