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. 2021 Jan 11;10(1):bio057539. doi: 10.1242/bio.057539

Fig. 3.

Fig. 3.

BPA-induced SG assembly requires PERK mediated phosphorylation of eIF2α. (A) Immunoblot detecting levels of phosphorylated eIF2α (top blot, labeled P-eIF2α) and total eIF2α (bottom blot) in U2OS cells treated as indicated with BPA, ARS (100 µM) for 1 h, or left untreated (indicated as ---). (B) Immunofluorescence detecting HuR (green), eIF4G (red), and Hoechst/DNA (blue) in wild-type MEFs co-cultured with U2OS cells (top panels), or MEFs with eIF2α-S51A point mutation co-cultured with U2OS cells (bottom panels), treated as indicated with BPA or ARS for 1 h or left untreated. MEFs exhibit punctate nuclear DNA. Scale bar indicates 10 microns. (C) Quantification of SGs in U2OS cells following siRNA-mediated knockdown of GCN2, HRI, PERK, and PKR, prior to 1 h BPA or ARS treatment, or no treatment. Graph represents mean with standard deviation; n≥3; * 0.05≥P.