Fig. 6: Reduced BMAL1 expression increased the ratio of P-eIF2α to total eIF2α and the number of stress granules.
From: Circadian control of stress granules by oscillating EIF2α
a–c Protein levels of p-eIF2α, eIF2α, and BMAL1 at indicated time points after stress shock in cells with reduced BMAL1 expression. Representative immunoblots are shown in a, and the quantification of p-eIF2α and eIF2α are presented in b and c, respectively (mean ± S.E.M.; n = 3 independent experiments, two-way analysis of variance (ANOVA) with Sidak’s multiple comparison test, * represents the P value of two-way ANOVA among 50–130 min ***P ≤ 0.001; # represents the P value of Sidak’s multiple comparison, #P ≤ 0.05). d, e Expression of total eIF2α blocked the increased stress granule formation by BMAL1 silencing. mCherry-eIF2α or mCherry was transfected in GFP-G3BP1 knock-in (KI) cells with control or small interfering RNA (siRNA) targeting BMAL1, and the number of stress granules formed was analyzed after stress shock. Representative images are shown in d, and the quantification is presented in e (mean ± S.E.M.; n = 150–200 cells per sample, *P ≤ 0.05, **P ≤ 0.01 by unpaired Student’s t-test between KD-BMAL1 and KD-BMAL1-eIF2α. Scale bar = 20 μm). Yellow arrows indicate mCherry and SG double-positive cells
