Fig. 2

Silencing of circ-ATXN1 affected the proliferation, migration and tube formation of GECs. a A schematic representation of how circ-ATXN1 arosed from the ATXN1 gene as determined by scanning ATXN1 genomic DNA and circBase. Sanger sequencing validated the sequence on the junction sites of circRNA-ATXN1. The black arrow indicated the head-to-tail splicing sites of circRNA-ATXN1. b The existence of circ-ATXN1 in GECs. c The expression of circ-ATXN1 in GECs treated with RNase R. Data represent mean ± SD (n = 3, each group; **P < 0.01, ^##P < 0.01). d The expression of line-ATXN1 in GECs treated with RNase R. Data represent mean ± SD (n = 3, each group; **P < 0.01, ^##P < 0.01). e The relative expression of circ-ATXN1 in AECs and GECs was detected by qRT-PCR. GAPDH was used as an endogenous control. Data represent mean ± SD (n = 3, each group; *P < 0.05) f The relative expression of line-ATXN1 was detected in AECs and GECs by qRT-PCR. GAPDH was used as an endogenous control. Data represent mean ± SD (n = 3, each group). g FISH was performed to investigate expression and location of circ-ATXN1 in AECs and GECs (green, circ-ATXN1; blue, DAPI nuclear staining). h Effect of circ-ATXN1 on the cell viability of GECs was detected by CCK-8 assay. Data represent mean ± SD (n = 3, each group; **P < 0.01). i Effect of circ-ATXN1 on the migration of GECs was detected by Transwell assay. Data represent mean ± SD (n = 3, each group; **P < 0.01). j Effect of circ-ATXN1 on tube formation of GECs was measured by Matrigel tube formation assay (Black arrow, tube structures; Grey arrow, tube branches). Data represent mean ± SD (n = 3, each group, **P < 0.01). Scale bar represents 30 μm. k-l Effect of circ-ATXN1 knockdown on expression of MMP2 and VEGFA was detected by Western blot analysis. Data represent mean ± SD (n = 3, each group; **P < 0.01). m The activity of MMP2 in the GECs after downregulation of circ-ATXN1 was detected by gelatin zymography. Data represent mean ± SD (n = 3, each group; *P < 0.05)