Figure 5.

ORF3a induced autophagy through unfolded protein response. (A) Hela cells (1–2 × 10⁵ cells) were transfected with HA-Tag-ORF3a (2 μg) or empty vector (2 μg) for 24 h. Western blot was performed to detect UPR-related proteins. (B) 293T cells (1–2 × 10⁵ cells) were transfected with HA-Tag-ORF3a (2 μg) or empty vector (2 μg) for 24 h pretreated with Ceapin-A7 (10 nM), GSK2606414 (5 nM), 4μ8C (10 nM) for 16 h before harvest. Western blot was performed to detect LC3B and p62. (C–E) Hela cells (1–2 × 10⁵ cells) were transfected with HA-Tag-ORF3a (2 μg) and atf6/perk/ire1 siRNA (50 nM) or control siRNA (50 nM) for 36 h. Western blot was performed to detect LC3B. One-way ANOVAs followed by Dunnett’s multiple comparisons test were used in comparing multiple groups. Student’s t-test was used to test statistical significance across two groups. Data are representative of 3 independent experiments. * p ≤ 0.05, **** p ≤ 0.001.