Figure S1.
Additional characterization of mtDNA-depleted IMR90 cells and expression of FAM43A in TFAM-depleted mouse liver tissue. (A) IMR90 cells were treated with ddC (20 µM) for the indicated time. After 96 h of treatment, cells were cultured in a ddC-free medium for 23 days. The relative mtDNA copy number of untreated (UNT) and mtDNA-depleted and repleted cells was measured by qPCR. Data points represent the mean of three technical replicates from three biological replicates. Error bars represent mean ± SD. (B) Representative immunoblot image of TFAM (bottom band) and p16 in UNT and ddC-treated IMR90 cells. ACTIN was blotted as a loading control. (C) Cumulative population doublings of IMR90 cells treated as described in A. Data points represent the mean of three technical replicates from three biological replicates. (D) Representative immunoblots of OXPHOS subunits in UNT and ddC-treated IMR90 cells. (E) Quantification of (D) (n = 4 biological replicates). Error bars represent mean ± SD, Unpaired Student’s t test; ns, P > 0.05; *, P < 0.05. (F) Transcription factor motif analysis of differentially expressed genes in cells treated with ddC (20 µM) for 96 h. False discovery rate (FDR) and normalized enrichment score are shown. (G) Relative mtDNA copy number of untreated (UNT) and IMR90 cells treated with ethidium bromide (EtBr) (50 ng/ml) for the indicated time measured by qPCR using ND1 primers normalized to nuclear B2M (n = 3 technical triplicates). Error bars represent mean ± SD, Unpaired Student’s t test; ****, P < 0.0001. (H) Relative FAM43A mRNA level of UNT and IMR90 cells treated with EtBr for the indicated time measured by RT-qPCR normalized to ACTIN (n = 3 technical replicates). Error bars represent mean ± SD, Unpaired Student’s t test; **, P < 0.01. (I) Relative expression of Fam43a and Tfam in liver samples isolated from Ctrl (WT) and Tfam+/flox C57BL/6 mice (n = 6 mice per condition). Error bars represent mean ± SD, Unpaired Student’s t test; **, P < 0.01; ***, P < 0.001. Source data are available for this figure: SourceData FS1.

Additional characterization of mtDNA-depleted IMR90 cells and expression of FAM43A in TFAM-depleted mouse liver tissue . (A) IMR90 cells were treated with ddC (20 µM) for the indicated time. After 96 h of treatment, cells were cultured in a ddC-free medium for 23 days. The relative mtDNA copy number of untreated (UNT) and mtDNA-depleted and repleted cells was measured by qPCR. Data points represent the mean of three technical replicates from three biological replicates. Error bars represent mean ± SD. (B) Representative immunoblot image of TFAM (bottom band) and p16 in UNT and ddC-treated IMR90 cells. ACTIN was blotted as a loading control. (C) Cumulative population doublings of IMR90 cells treated as described in A. Data points represent the mean of three technical replicates from three biological replicates. (D) Representative immunoblots of OXPHOS subunits in UNT and ddC-treated IMR90 cells. (E) Quantification of (D) (n = 4 biological replicates). Error bars represent mean ± SD, Unpaired Student’s t test; ns, P > 0.05; *, P < 0.05. (F) Transcription factor motif analysis of differentially expressed genes in cells treated with ddC (20 µM) for 96 h. False discovery rate (FDR) and normalized enrichment score are shown. (G) Relative mtDNA copy number of untreated (UNT) and IMR90 cells treated with ethidium bromide (EtBr) (50 ng/ml) for the indicated time measured by qPCR using ND1 primers normalized to nuclear B2M (n = 3 technical triplicates). Error bars represent mean ± SD, Unpaired Student’s t test; ****, P < 0.0001. (H) Relative FAM43A mRNA level of UNT and IMR90 cells treated with EtBr for the indicated time measured by RT-qPCR normalized to ACTIN (n = 3 technical replicates). Error bars represent mean ± SD, Unpaired Student’s t test; **, P < 0.01. (I) Relative expression of Fam43a and Tfam in liver samples isolated from Ctrl (WT) and Tfam+/flox C57BL/6 mice (n = 6 mice per condition). Error bars represent mean ± SD, Unpaired Student’s t test; **, P < 0.01; ***, P < 0.001. Source data are available for this figure: SourceData FS1.

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