Supplementary Figure 2: CL conditions for regulation of mitochondrial morphology and quality control.
From: Molecular basis of selective mitochondrial fusion by heterotypic action between OPA1 and cardiolipin
(a) Comparison of the number, length, and unsaturation of acyl chains of CL used in this study. (b) GTP hydrolysis activity of the deletion mutant (Δ581–941) of L-OPA1 (n = 3 independent measurements, data are mean ± s.d.). (c–g) KD of CLS1 using siRNA. (c) RNAs were analyzed by RT-PCR (the experiment was performed one time). Nuclear genome-encoded transcripts were analyzed. (d) Fluorescence images of the control and CLS1 KD cells (representative images of more than 3 independent experiments with similar results). Scale bar: 10 μm. (e) Immunoblot analysis of mitochondrial protein levels using the indicated antibodies (the experiment was performed one time). (f,g) Mitochondrial fusion in CLS1 KD cells and OPA1 KD cells. Mitochondrial fusion was analyzed using HeLa cells expressing mitGFP and mitRFP that were co-cultured and fused with the HVJ envelope. After 6 h culture, fusion efficiency was analyzed by fluorescence microscopy. n, number of cell hybrids analyzed from 3 independent experiments: cont × cont (n = 38), CLS1KD × CLS1KD (n = 41), and OPA1KD × OPA1KD (n = 32). Scale bar: 10 μm. Statistics source data are available in Supplementary Table 1. Unprocessed original scans of agarose gel and blots are shown in Supplementary Fig. 3. (h) Model of the isolation of damaged mitochondria from the active mitochondrial network. OPA1 inactivation and CL remodeling are used as markers for the specific selection of damaged mitochondria.
