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. 2022 Apr 4;10:871977. doi: 10.3389/fcell.2022.871977

FIGURE 3.

FIGURE 3

vMIA-induced peroxisomal and mitochondrial fragmentation is independent of MAVS. vMIA does not disrupt STING-MAVS interaction at peroxisomes. (A) Immunofluorescence analyses of MEFs MAVS KO cells: (a, b) peroxisomal and mitochondrial morphologies in control cells: (a) anti-PMP70, (b) anti-TIM23; (c–e) peroxisomal morphology upon overexpression of vMIA-Myc: (c) anti-PMP70, (d) anti-Myc, (e) merge image of c and d; (f, h) mitochondrial morphology upon overexpression of vMIA-Myc: (f) anti-TIM23, (g) anti-Myc, (h) merge image of f and g. Bars represent 10 µm. (B,C) Statistical analysis of peroxisomal or mitochondrial morphologies upon overexpression of vMIA-Myc in MEFs MAVS KO cells, respectively. Approximately 600 cells were analysed per condition. (D) Co-immunoprecipitation analysis of the interaction between overexpressed STING-FLAG and vMIA-Myc in MEFs MAVS-PEX cells. The pull-down was performed using an antibody against MAVS. Western blot was performed with antibodies against FLAG and Myc. IN represents total cell lysate (input), IP represents immunoprecipitation and OUT represents the cell lysate extracted after incubation with the antibody (output). (E) Quantification of the ratio between IP and IN, in the presence or absence of vMIA. Data represents the means ± SEM of three independent experiments, analysed using unpaired T test (ns - non-significant; ***–p < 0.001, ****–p < 0.0001).