Supplementary Figure 9 from Aberrant CircTMEM45A Facilitates Inflammatory Progression of Esophageal Squamous Cell Carcinoma through m5C-Mediated NLRP3 Activation

Supplementary Figure 9. Interactions between circTMEM45A and NSUN2, ALYREF, and YBX1 proteins. (A-B) qRT-PCR and western blot analyses of NSUN2, ALYREF, and YBX1 mRNA and protein expression in circTMEM45A-overexpressed KYSE30 cells and circTMEM45A-silenced KYSE150 cells. (C-H) qRT-PCR and western blot verified the knockdown efficiency of NSUN2 (C-D), ALYREF (E-F) and YBX1 (G-H) in KYSE150 cells. (I) qRT-PCR analysis of circTMEM45A expression following the knockdown of NSUN2, ALYREF, and YBX1 in KYSE150 cells. (J) RIP assay demonstrating the enrichment of NLRP3 by ALYREF (Left) and YBX1 (Middle) antibodies in KYSE150 cells; Right, RAP assay showing the enrichment of the circTMEM45A probe for NLRP3 in KYSE150 cells. (K) qRT-PCR analysis of circTMEM45A subcellular localization after knocking down NSUN2 or ALYREF in KYSE150 cells. The relative gray value (target/GAPDH) is the average of all independent repeated experimental data (n=3). ns: not significant, ***P<0.001.