Fig. 5

TRPM2-AS promotes the activation of NOTCH1 signaling pathway in HUVECs. A, B Transcriptomic sequencing was performed on 1 × 10⁷ control and TRPM2-AS overexpressing HUVECs cells, and KEGG was performed to analyse the significantly altered signaling pathways. C-E Western blot validation of HEY1/HES1/N1ICD/NOTCH1 expression level in TRPM2-AS overexpressing, knockdown, and control HUVECs and immunofluorescence verification of N1ICD expression level in TRPM2-AS overexpressing, knockdown, and control HUVECs and in HUVECs cultured in Nc-exo/Ts-exo with/without knockdown of TRPM2-AS. Green fluorescence: corresponding primary antibody stained N1ICD; blue fluorescence: DAPI-stained nuclei. Scale bar: 20 μm. F, G Western blot analysis showing the HEY1/HES1/N1ICD/NOTCH1 expression levels and representative immunofluorescence images of N1ICD expression in TRPM2-AS overexpressing/control HUVECs with/without DAPT treatment and in HUVECs cultured in Nc-exo/Ts-exo with/without DAPT treatment. H, I EDU, transwell, and tube formation assays to detect the angiogenic ability of TRPM2-AS overexpressing/control HUVECs with/without DAPT treatment (H), and HUVECs cultured in Nc-exo/Ts-exo with/without DAPT treatment (I). Red fluorescence: EDU-positive cells (EDU⁺). Scale bar: 200 μm (EDU/transwell/tube formation). Data were assessed with unpaired Student’s t test or one-way ANOVA and presented as mean ± SD. * P < 0.05; ** P < 0.01; *** P < 0.001; ns, no significance