CHX treatment induces HPIV3-triggered SG disassembly.

(A and B) HeLa cells were treated with AS (0.5 mM) for 30 min and subsequently treated with or without CHX in the presence of AS for another 1 h. (A) Cells were immunostained for TIA-1 (green) and G3BP (red). Nuclei were stained with DAPI (blue). The white scale bar corresponds to 10 μm. (B) The percentage of cells containing SGs was quantified in three independent experiments. (C-H) HeLa cells were mock-infected or infected with HPIV3 (MOI = 1) for 24 h and subsequently mock-treated or treated with CHX for another 1 h, 2 h, or 3 h. (C) Cells were immnostained for HPIV3 (purple), TIA-1 (green), and G3BP (red). Nuclei were stained with DAPI (blue). The white scale bar corresponds to 10 μm. (D) The percentage of cells containing SGs was quantified in three independent experiments. (E) Cell lysates were analyzed via western blot using anti-HN and anti-GAPDH antibodies. (F) Western blots from three independent experiments were quantified and normalized with respect to the amount of GAPDH. (G and H) Total RNA were isolated for qPCR analysis to measure the indicated RNA abundance and normalized to that of GAPDH. Data are represented as means ±SD. Student’s t test: * P<0.05, ** P<0.01, *** P<0.001, ns = not significant.