Fig. 4

circRHOBTB3 inhibits EMT by binding to metabolism-related proteins to regulate ROS. (A) GSEA and (B) ROS levels of circRHOBTB3-KO SW480 cells compared with mock SW480 cells. (C) Western blot of mock and circRHOBTB3-KO SW480 cells using anti-E-cadherin and anti-vimentin antibodies. (D) Cell migration, invasion, and (E) proliferation assays and (F) Western blot of control and LNAC-treated circRHOBTB3-KO SW480 cells. The right column diagram shows the quantification analysis results. (G) Schematic diagram of biotin-labeled circular RNA synthesis. (H) Verification of the synthesized biotin-labeled circRHOBTB3. (I) Different protein bands detected by silver staining for mass spectrometry. (J) Western blot of RNA pull-down products using anti-Eno1 and anti-Eno2 antibodies. Anti-GAPDH were used as negative control. (K) ClueGO functional analysis of potential proteins interacting with circRHOBTB3. All experiments were repeated for three times, data were shown as mean ± SD (E) or mean ± SEM (B, D), * P < 0.05, ** P < 0.01, *** P < 0.001, NS P > 0.05, in Student’s test (E) or paired Student’s test (B, D)