Fig. 1

CALCOCO2 and SQSTM1 differentially regulate CVB3 propagation. a HeLa cells were transiently transfected with siRNAs targeting CALCOCO2 (siCALCOCO2) or SQSTM1 (siSQSTM1), or a scramble siRNA control (siCON) for 48 h, followed by CVB3 infection (MOI = 0.1) for 24 h. Western blotting was performed to examine protein expression of CALCOCO2, SQSTM1, VP1, and ACTB. Protein levels of VP1 were quantified by densitometric analysis using NIH ImageJ, normalized to ACTB and presented underneath as fold changes compared to sham (the first lane of sham is arbitrarily set a value of 1). b HeLa cells were treated as above. Cell-associated virus titers were determined by TCID₅₀. Data are represented as mean ± SD from three replicates. c, d HeLa cells were treated with siCALCOCO2 as above, and then subjected to infection with different doses of CVB3 for various times as indicated. Western blotting and densitometric analysis were conducted (c) and virus titers (mean ± SD, n = 3) were measured (d) as above. e–h HeLa cells were transfected with constructs overexpressing Flag-CALCOCO2 (e, f) or Flag-SQSTM1 (g, h) for 24 h and then subjected to CVB3 infection (MOI = 0.1) for 16 h. Cell lysates were analyzed by western blotting for VP1 protein levels (e, g) and cell-associated virus titers were quantified as above (f, h). Results in this figure represent data from two to three independent experiments