Figure 4.

Transient knockdown of syntaxin-17 (STX17), synaptosome-associated protein of 29 kDa (SNAP29), LC3B, and lysosomal-associated membrane protein 1 (LAMP1) impairs production of infectious EV-A71. (A) RD cells were transfected with 40 nM of STX17, SNAP29, LC3B, LAMP1, or control pools of 3 siRNAs for 48 h, and endogenous proteins were detected by Western Blot (WB); (B) RD cells were transfected as in (A) and infected with EV-A71 (MOI = 10) for 8 hpi, and SQSTM1/p62 and viral proteins were detected by WB; (C) RD cells were transfected and infected as in (B) for plaque assay to determine viral titres; (D) Graphical presentation of the 47 autophagy-associated proteins considered in this study. These proteins were used in the yeast two-hybrid arrays against 2BC non-structural protein; (E) Graphical presentation of the 12 autophagy-associated proteins that interacted with 2BC non-structural protein of EV-A71. The Markov Cluster Algorithm was performed using STRING to cluster the autophagy-related proteins. The width of blue lines is proportional to the strength of interactions between proteins, with greater width indicating stronger interaction. The dashed lines indicate the possible interactions between proteins. One-way ANOVA: *** p < 0.001, and **** p < 0.0001.