Extended Data Fig. 6: Expression of HIF1A and VHL in clonal HAP1 lines with variants assayed in SGE.
From: Saturation genome editing maps the functional spectrum of pathogenic VHL alleles
a,b, Clonal HAP1 cell lines were isolated containing SNVs introduced independently via prime editing. Western blots were performed to assess VHL and HIF1A protein levels, with α-tubulin stained as a loading control. SNVs scored as significantly depleted in SGE (all except c.228C>G, c.222C>A and c.191G>C) showed increased levels of HIF1A expression compared to unedited HAP1 and cells expressing p.F76L, a variant scored neutrally by SGE. Of note, c.222C>A and c.462A>C had RNA scores of −4.45 and −5.82, respectively. Clonal variability may account for subtle differences between results from the SGE assay and the degree of HIF1A upregulation observed by western blot. (Results are representative of 2 independent blots per line.)