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. 2023 Feb 20;14(2):141. doi: 10.1038/s41419-023-05678-3

Fig. 5. TIM-4 interacts with ANXA2 to regulate the PI3K/AKT signaling pathway and mitochondrial function.

Fig. 5

A Venn diagram depicting the comparison of proteins identified in A549-LV-CON, A549-LV-TIM-4, and HEK-293 cells. The shared proteins were excluded from the differential lists. B, C Co-immunoprecipitation (Co-IP) assays were performed in A549 and H23 cells overexpressing TIM-4 or control. D, E OCR, mitochondrial basal respiration and maximal respiration of A549 cells and H23 cells transfected with siRNA of ANXA2 or control. F Mitochondrial membrane potential (JC-1 aggregates) in A549 cells transfected with siRNA targeting ANXA2 were accessed by the JC-1 assay kit according to the manufacture’s instruction. G Mitochondria fitness was tested with Mito-tracker Deep Red. H Flow cytometry dot plots showed the percentage of depolarized mitochondria in A549 cell lines transfected with siRNA targeting ANXA2. I Western blotting analysis of OPA1, p-AKT, PCNA, and ANXA2 expression in siANXA2-transfected lung cancer cell lines. J Flow cytometry histograms showing the level of Ki67 in lung cancer cell lines transfected with siRNA targeting ANXA2. Three independent experiments were conducted for each result and error bars represent SEM per group in one experiment. Data were analyzed using Student’s t test (two-tailed unpaired t test) for (D), (E), (F–H), and (J). ns means non-significance; *P < 0.05; **P < 0.01; ***P < 0.001.