Figure 5. Effect of proton flux on NF-κB activation and Jmjd3 expression in methyl-β-cyclodextrin (MCD)-treated cells.

(A) Schematic of potential mechanism induced by MCD, and MCD/carbonyl cyanide m-chlorophenyl hydrazine (CCCP) on mitochondrial proton flux. (B) Effect of CCCP on NF-κB activity and Jmjd3 expression: (a) on NF-κB activity in RAW blue macrophages (CCCP = 50 µM); (b) effect of CCCP on Il1b expression in RAW 264.7 macrophages (CCCP = 50 µM). (C) Effect of CCCP (a) on Jmjd3 expression in RAW 264.7 macrophages (CCCP = 50 µM) and (b) on bone marrow-derived macrophages (BMDMs) (CCCP = 200 µM). Data are representative of 3 independent experiments with 3 samples per group and data are presented as mean ± standard deviation (SD). Statistical analysis was performed using unpaired, two-tailed Student’s t-test. Asterisks (*) and (**) indicate a significant difference with p < 0.05 and p < 0.001. A hashtag (#) indicates a significant difference between MCD without or with inhibitors with p < 0.05.

Figure 5—figure supplement 1. The effect of proton flux Inhibitors.

(A) The toxicity of carbonyl cyanide m-chlorophenyl hydrazine (CCCP) by 3- (4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. (B) Effect of BAM15 (200 µM) on Il1b (a) and Jmjd3 (b) gene expression in methyl-β-cyclodextrin (MCD)-treated RAW264.7 macrophages. Data are representative of 3 independent samples per condition and are presented as mean ± standard deviation (SD). Statistical analysis was performed using unpaired, two-tailed Student’s t-test. Asterisk (*) and hashtag (#) indicate a significant difference between MCD without or with inhibitors with p < 0.05; (**) indicates a significant difference with p < 0.01.