Fig. 3

Induction of EndMT by MFNG overexpression in HUVECs. a qRT-PCR analysis was conducted to determine the relative mRNA levels of MFNG, with GAPDH as the loading control. b Western blot confirmed MFNG overexpression in HUVECs, with β-actin as the loading control. The band density of MFNG was quantified. c Light microscopy (upper) illustrated the morphological changes in HUVECs after MFNG overexpression. F-actin staining with phalloidin using a fluorescence microscope (lower) revealed the transformation of HUVECs into mesenchymal stellate cells with long protrusions and filopodia 48 h post-transfection. Scale bars = 25 µm. d Western blot depicted alterations in EndMT-related markers in HUVECs with MFNG overexpression compared to the control group, with a quantitative analysis of the relative protein expressions of MFNG, CD31, SLUG, VIMENTIN, and α-SMA using ImageJ software. e Cellular immunofluorescence staining for CD31 (red), VIMENTIN (red), and MFNG (green). Nuclei were counterstained with DAPI. Scale bars = 25 µm. f, g Representative images of cell wound healing assays and quantification of the scratch repair rate. Scale bars = 250 µm. h, i Assessment of the migratory ability of HUVECs overexpressing MFNG via the transwell assay and quantification of migrated HUVECs. Scale bars = 100 µm. All data presented are from three independently repeated biological experiments