Fig. 4: DAG enhances Drp1-mediated fission of OMM-mimetic templates. | Nature Communications

Fig. 4: DAG enhances Drp1-mediated fission of OMM-mimetic templates.

From: Acute diacylglycerol production activates critical membrane-shaping proteins leading to mitochondrial tubulation and fission

Fig. 4

a Plots showing the fraction of membrane tubes with at least one cut in the presence of Drp1 and GTP. Data represents the analysis of at least 120 tubes for each of the indicated SMrT preparations. Membrane tubes contained either 5% DAG or 5% PI in a background of 5% CL (none). b Representative images of membrane tubes of the indicated lipid composition after 10 mins incubation with Drp1 and GTP. c Representative images (top) and fluorescence line profiles (bottom) of a membrane tube (magenta) with GppNHp-bound Drp1-mEGFP scaffolds (green). d The membrane density of Drp1-mEGFP scaffolds is reported as the ratio of the EGFP fluorescence and the membrane fluorescence on membrane tubes representing a range of sizes prepared using the indicated lipid composition. Data is normalized to the highest mean density for each condition and represents the mean \(\pm \,\)SD of at least 47 tubes. Statistical significance was estimated using unpaired Mann-Whitney’s test. **** denotes p < 0.0001, *** denotes p = 0.001, ** denotes p = 0.002, * denotes p = 0.02. e Representative images showing GppNHp-bound Drp1-mEGFP (green) on membrane tubes (magenta) displaying the 6xHis-tagged soluble domain of mitochondrial fission factor (MFF), which is recruited to the membrane by incorporating 5% of the DGS-NTA(Ni2+) lipid into templates. f The membrane density of Drp1-mEGFP is reported as the ratio of the EGFP fluorescence and the membrane fluorescence on membrane tubes representing a range of sizes prepared using the indicated lipid composition. Data is normalized to the highest mean density and represents the mean \(\pm \,\)SD of at least 30 tubes for each condition. Statistical significance was estimated using unpaired Mann-Whitney’s test. **** denotes p < 0.0001, *** denotes p = 0.001, ** denotes p = 0.009, * denotes p = 0.03. g Representative images of MFF- and CL-containing tubes (gray) before and after 30 mins incubation with Drp1 and GTP. h Quantification of the frequency of cuts per membrane tube for MFF-containing templates prepared with 5% PI or 5% DAG. Data represents the analysis of at least 169 tubes for each condition.

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