Fig. 4: miR-1224-5p regulates the autophagic degradation of EGFR and downstream EFNA1/EPHA2 and VEGFA signalling pathways.

a–d qRT-PCR and western blotting assays were used to examine the mRNA and protein levels of EGFR in ESCC cells transfected with miR-1224-5p mimic or inhibitor. e, f Western blotting assays were performed to measure EGFR expression in miR-1224-5p mimic-transfected and BafA1 (100 nM)-treated KYSE150 and KYSE510 cells. g, h A co-IP assay was used to examine the interaction between TNS4 and EGFR in KYSE150 and KYSE510 cells. i, j Western blotting was used to measure the protein levels of EGFR in TNS4-silenced KYSE150 and KYSE510 cells. k–r In miR-1224-5p-overexpressing or miR-1224-5p-silenced ESCC cells, qRT-PCR, western blotting and ELISA assays were used to examine the mRNA level of EFNA1 and the protein levels of EFNA1, p-EPHA2, EPHA2, VEGFA and VEGFA in the ESCC cell supernatant. s, t Western blotting and ELISA assays were performed to measure the protein levels of EGFR, EFNA1, VEGFA and VEGFA in the ESCC cell supernatant after EGFR knockdown. Data are represented as the mean ± SD, and experiments were performed in triplicate; * p < 0.05, ** p < 0.01, *** p < 0.001, ns not significant.