Figure 7.

Mitophagy has an effect on the LDHB-mediated NFKB signaling pathway activation. (A) PK-15 and 3D4/2 cells were treated with DMSO or CCCP (10 uM) for 2 h, then mock-inhibited or inhibited with the siRNA of LDHB. Western blot analysis of the relative expression of the mitophagy mark proteins of TOMM20 and VDAC1, the NFKB signal pathway protein IKBIA, RELA, NKIRAS2, and TUBA was as a loading control. The level of protein was quantified using Image-Pro Plus 6.0 software. Error bars indicate the mean (±SD) of 3 independent experiments. *, P < 0.05; **, P < 0.01; and ***, P < 0.001 (one-way ANOVA). (B) PK-15 and 3D4/2 cells were transfected with shRNA of PRKN or non-targeting shRNA for 24 h, then mock-inhibited or inhibited with the siRNA of LDHB. Western blot analysis of the relative expression of PRKN protein to detect the effective knockdown of shPRKN, the expression of TOMM20 and VDAC1, the NFKB signal pathway protein IKBIA, RELA, NKIRAS2, the loading control protein TUBA were detected as in Figure 6A. (C) q-RT-PCR was used to detect the relative expression of cytokines IFNA, IFNB, and TNF as described in Materials and Methods. Error bars indicate the mean (±SD) of 3 independent experiments. *, P < 0.05; **, P < 0.01; and ***, P < 0.001 (one-way ANOVA)