Fig. 7

Cleaved PGAM5 enhances mitochondrial biogenesis A Mitochondria were visualized by MitoTracker Red staining in hippocampal neurons. Scale bar, 20 µm. B Quantification of total MitoTracker intensity in individual neurons. Dashed lines indicate the medium and dotted lines indicate the 25th and the 75th percentiles (n = 181–190 cells/group). * p < 0.05, ** p < 0.01, Student’s t-test. C Immunoblot of PGC1α, NRF1, TFAM and TIM23 in control and injured cortical neurons. D, E Quantification of (C) on DIV9 and DIV10 respectively. Data are presented as mean ± SEM (n = 6). * p < 0.05, Student’s t-test. F Immunoblot of PGC1α, PGAM5 and TIM23 in neuro2a cells overexpressing full-length PGAM5, PGAM5(Δ2-24) or full-length PGAM5 + hPARL(DDD). G Percentage of cleaved PGAM5 in neuro2a cells overexpressing full-length PGAM5 with or without co-expressing hPARL(DDD). Data are presented as mean ± SEM (n = 5). * p < 0.05, paired t-test. H, I Quantification of (F). Data are presented as mean ± SEM (n = 5). * p < 0.05, one‑way ANOVA with Dunnett’s multiple comparisons. J Immunoblot of TFAM in brain tissue lysate of CCI mice on 4 dpi. K Quantification of relative TFAM in J. Data are presented as mean ± SEM (n = 6). * p < 0.05, one‑way ANOVA with Tukey’s multiple comparisons. L Immunoblot of PGAM5 and TFAM in neuro2a cells transiently transfected with shlacZ or shPgam5 vector. M, N Quantification of relative PGAM5 and TFAM in (L). Data are presented as mean ± SEM (n = 6). * p < 0.05, *** p < 0.001, paired t-test. O Images of neuro2a cells overexpressing EGFP, PGAM5-GFP, PGAM5(Δ2-24)-GFP or PGAM5-GFP + hPARL(DDD). Mitochondria were visualized by MitoBright LT Deep Red. Scale bar, 20 μm. P Quantification of total intensity of MitoBright LT Deep Red in individual cells. Dashed lines indicate the medium and dotted lines indicate the 25th and the 75th percentiles (n = 188–194 cells/group). * p < 0.05, ** p < 0.01, **** p < 0.0001, one‑way ANOVA with Tukey’s multiple comparisons