Time-series RNA-seq of U2OS cells following CCCP-induced mitochondrial damage and recovery upon drug washout

Mitochondria are continuously exposed to damage that contributes to aging and disease. While prolonged mitochondrial damage triggers mitophagy, how cells respond to transient damage remains unclear. To investigate the transcriptional programs underlying transient mitochondrial damage and subsequent recovery, we performed time-series RNA-seq analysis in human U2OS cells treated with the mitochondrial uncoupler CCCP. Four groups were analyzed: untreated control (WT), 50 μM CCCP for 30 min (CCCP), 50 μM CCCP for 30 min followed by 1 h washout (W1H), 50 μM CCCP for 30 min followed by 6 h washout (W6H). Each condition was analyzed in triplicate. This dataset reveals the dynamic transcriptional landscape during mitochondrial damage and recovery, highlighting the activation of stress-responsive transcription factors and antioxidant defense pathways that facilitate mitochondrial repair.