Figure 6.

RRV-induced phosphorylation of eIF2α is not blocked by G0. (A) Heatmap showing deregulation of genes associated with EIF2 signaling pathway in T-REx-293 cells expressing APOL1 RRVs. Western blot analysis of phosphorylation level of eIF2α (S51) from cell lysates of T-REx-293 (B) monoallelic and (C) biallelic clones after 8 hours of tet induction: no treatment, intermediate (12.5 or 25 ng/ml), or high (50 ng/ml) tet dose]. (D) Measurement of protein synthesis rate. The incorporation of puromycin into newly synthesized proteins was detected by immunoblotting with anti-puromycin antibody. Bar diagram showing the level of nascent proteins (normalized with internal control, β-actin) in T-REx-293 cell expressing increasing level of APOL1 variants after 8 hours of tet induction (no treatment, intermediate [12.5/25 ng/ml], or high [50 ng/ml] tet dose) and 10 minutes of incubation with 10 μg/ml puromycin.