Fig. 5: KML001-induced telomeric DNA damage activates p53-mediated repression of the PGC-1α gene network and mitochondrial functions.

A Kinetic analysis of p53, γH2AX, PARP1 protein levels by western blot in CD4 T cells exposed to KML001 or DPBS control for 6, 12, 24, and 48 h. B Representative western blots for the key molecules in the PGC-1α network in CD4 T cells treated with DPBS control, or 5 or 10 μM KML0001 for 48 h in the presence of anti-CD3/CD28 antibody stimulation. C Representative western blot bands for the PGC-1α network following TP53 knockdown (KD) by CRISPR/Cas9 nucleofection. D Summary data of RT-qPCR analysis of TP53 gene expression following siRNA-mediated TP53 KD (n = 7). E Representative western blot bands for the PGC-1α network following TP53 KD by siRNA. F Summary data for the frequency of apoptotic Av⁺ cells in CD4 T cells following TP53 KD (n = 10). G Summary data for the MFI of MG in CD4 T cells following TP53 KD (n = 4). H The CD4 T cell oxygen consumption rate (OCR) was measured by Seahorse XFp Cell Mito Stress Tests, and summary data for basal respiration, maximal respiration, spare respiratory capacity, and ATP production following TP53 KD (n = 6) are shown. I ATP production measured by a luminescence test in CD4 T cells following p53 KD (n = 6). J Mitochondrial DNA (mtDNA) relative to nuclear DNA (nuDNA) content was analyzed by qPCR following TP53 KD (n = 8). K Representative western blot bands and summary data following TRF2 KD (n = 5). L The oxygen consumption rate (OCR) was measured by Seahorse XFp Cell Mito Stress Tests, and summary data for basal respiration, maximal respiration, spare respiratory capacity, and ATP production following TRF2 KD (n = 5) are shown. M A model depicting the molecular mechanism linking telomeric DNA damage and mitochondrial compromise via KML001-mediated telomere shortening and p53-mediated repression of the PGC-1α gene network. All summary data are shown as mean ± SEM.