Figure 5.

IRE1α-JNK signaling pathway is involved in DENV2-induced autophagy and affects viral titer. (A) Huh7 cells were infected with DENV2 (MOI = 10) and the cells were collected at 36 h p.i. (B) Huh7 cells were transfected with sh-IRE1α for 18 h and infected with DENV2 (MOI = 10). (C) Huh7 cells were infected with DENV2 (MOI = 10) followed by SP600125 treatment at different concentrations. (D) Huh7 cells were infected with DENV2 then treated with or without SP600125 (30 μM), and the cells were fixed at 36 h p.i. The FITC-labeled DENV2 NS1 (green), PE-labeled LC3 puncta (red) and Hochest labeled nuclei (blue) were detected under the fluorescent microscopy. Arrow head points the LC3 puncta. The protein levels of IRE1α, p-JNK, JNK, p-Bcl-2, Bcl-2, LC3-II/ LC3-I and NS1 were determined by Western blotting at 36 h p.i. The number under the band is the quantification of band intensity after normalization with β-actin. The viral titer was determined by plaque assay.