Skip to main content
. 2022 Jun 28;54:102388. doi: 10.1016/j.redox.2022.102388

Fig. 6.

Fig. 6

Small molecule reducing agents promotes ORF8 degradation and mitigates ER stress.

(A, B) Immunoblotting of the UPR markers in PDI KO (A) or ERP44 KO (B) HepG2 cells expressing ORF8-FLAG.

(C) Immunoblotting analysis of the redox state of ORF8 in HEK 293T cells expressing ORF8-FLAG treated with 1 mM NAC or VC for 24 h, or 200 μM DTT for 6 h. Cell lysates were separated by SDS-PAGE under either reducing (R) or nonreducing (NR) conditions.

(D) Immunoblotting of ORF8 in HEK 293T cells expressing ORF8-FLAG treated with 1 mM NAC, 1 mM VC or 200 μM DTT for 15 h and then followed by treatment with 25 μg/ml CHX for indicated time points.

(E) The relative ratios of ORF8/GAPDH normalized to that at zero time point were quantified. The band intensities of ORF8 and GAPDH in (D) were analyzed using the ImageJ software. The data were shown as mean ± SEM from three independent experiments; *p < 0.05 and ***p < 0.001 (two-tailed Student's t-test).

(F) Detection of the UPR markers by immunoblotting in HEK 293T cells expressing ORF8-FLAG treated with 1 mM NAC or VC for 24 h, or 200 μM DTT for 6 h.