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. 2020 Apr 9;8:180. doi: 10.3389/fcell.2020.00180

FIGURE 4.

FIGURE 4

OPA1-Exon4b modulates mtDNA transcription. (A,B) Anti-Flag ChIP was carried out using WT MEF cells expressing Flag and Opa1 KO cells expressing Flag, OPA1-iso1-Flag, or OPA1-iso5-Flag. The precipitated DNA was analyzed by qPCR using primer pairs for the D-loop region (A) or Cox1 (B). n = 3, biological replicates. (C) Anti-Flag ChIP was carried out using WT MEF cells expressing Flag and Opa1 KO cells expressing Flag, NT-Exon4-Flag, or NT-Exon4/4b-Flag. The precipitated DNA was analyzed by qPCR using primer pairs for the D-loop region. n = 3, biological replicates. (D) Relative mRNA levels of 9 mtDNA genes in WT MEF cells expressing Flag and Opa1 KO cells expressing Flag, OPA1-iso1-Flag, or OPA1-iso5-Flag (n = 3, biological replicates). (E) Relative mRNA levels of three nuclear genes encoding respiratory subunits in WT MEF cells expressing Flag and Opa1 KO cells expressing Flag, OPA1-iso1-Flag or OPA1-iso5-Flag (n = 3, biological replicates). (F) Western blotting analysis of mtDNA-encoded Cox2 in WT MEF cells expressing Flag and Opa1 KO cells expressing Flag, OPA1-iso1-Flag or OPA1-iso5-Flag. Band densities were quantified using ImageJ, and relative band densities are shown on the bottom. n = 3, biological replicates. *p < 0.05, **p < 0.01, one-way ANOVA.