Fig. 5: circATXN1 Inhibits Partial EndMT by Suppressing SLUG.
From: Regulation of partial endothelial-to-mesenchymal transition by circATXN1 in ischemic diseases
a Western blot of SLUG, SNAIL, ZEB1, ZEB2, and TWIST in HCMECs with Ad Scr-sh or Ad circATXN1-sh for 48 h (n = 3 biological replicates). b FUCCI images of HCMECs infected with Ad Scr-sh, Ad Scr-sh + Ad SLUG-sh, Ad circATXN1-sh, or Ad circATXN1-sh + Ad SLUG-sh, and hypoxia-stimulated for 24 h (n = 3 biological replicates, scale bars = 100 µm). c, d Transwell and spheroid sprouting assays of HCMECs infected as above, with hypoxia stimulation for 24 h (n = 4 biological replicates for c, scale bars = 100 µm. n = 6 biological replicates for d, scale bars = 100 µm). e Western blot of Fibronectin, VE-cadherin, CD31, αSMA, and FSP1 in HCMECs infected and stimulated as above (n = 3 biological replicates). f RT-qPCR of SLUG in HCMECs infected with Ad Control or Ad circATXN1 (n = 3 biological replicates). g Dual luciferase reporter assay of SLUG promoter activity in HCMECs infected with Ad Control or Ad circATXN1 (n = 5 biological replicates). h Western blot SLUG in HCMECs infected with Ad Control + Ad Scr-sh, Ad circATXN1 + Ad Scr-sh, Ad Control + Ad AGO2-sh, or Ad circATXN1 + Ad AGO2-sh (n = 3 biological replicates). i RIP-qPCR analysis of circATXN1 and circHIPK3 in HCMECs with anti-IgG or anti-AGO2 (n = 3 biological replicates). j Western blot of mCherry expression in HEK293T cells transfected with Control, circATXN1-peptide1-mCherry, circATXN1-peptide2-mCherry, or mCherry for 48 h. β-actin served as the internal control (n = 3 biological replicates). Statistical analysis: ordinary one-Way ANOVA test for (c, d); unpaired two-tailed Student t test with Welch’s correction for (f); unpaired two-tailed Student t test for (g); two-Way ANOVA with multiple comparisons test for (i).
