Fig 2: Representative immunofluorescence staining for RNA methylation regulators (green) and DAPI (blue) in different groups of human podocytes. (A) Immunofluorescence co-localization was used to analysis the expression of METTL3, ADAR1 (B), DNMT1 (C) and WTAP (D). Scale bars: 10 μm. HG, high glucose, 30 mM; NG, normal glucose, 5.5 mM

Fig 3: Validation of functional phenotypes in database and cell lines. (A) Correlations between five RNA methylation regulatory genes and eGFR. (B) Relative expression levels of METTL3, WTAP, YTHDC1, ADAR1, DNMT1 mRNA in immortalized human podocyte cell line (HPC) and mouse podocyte cell line (MPC) by qRT-PCR. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001 versus normal glucose (NG) group. (C, D) Immunoblot analysis of RNA methylation regulators protein in high glucose (HG) induced podocytes. (E, F) Immunoblot analysis of podocyte damage-related protein in high glucose (HG) induced podocytes.β-actin served as a loading control

Fig 4: ADAR1 directs CYP1A1 A-to-I RNA editing. Relative expression of ADAR1 (A) and ADAR2 (B) between NSCLC tissues (Tumor) and matched normal adjacent tissues (NAT), and normalized expression levels of target genes were calculated relative to GAPDH using the ΔΔCT method (Wilcoxon signed rank test). C Western blot showing expression of ADAR1 and ADAR2 proteins in NSCLC tissues (T) and matched normal adjacent tissues (N) specimens from individuals 28, 33, 69, 71, and 96. GAPDH was the loading control. Correlations between CYP1A1 editing levels and the relative normalized mRNA levels of ADAR1 (D) and ADAR2 (E) of 34 paired specimens. Solid lines represent the linear regression (r), and dashed lines represent the 95% confidence interval (Spearman correlation coefficient test). F Western blot showing the expression of ADAR1 and ADAR2 proteins in A549 cells transiently transfected with the indicated expression constructs. CTL, control construct. β-Actin was the loading control. G, H, J, K Sequence chromatograms of the CYP1A1 transcript in the indicated cell lines; the arrowheads indicate the edited positions. (I) Western blot of ADAR1 and ADAR2 in 95D cells transiently transfected with expression plasmids as indicated. sh-NC, negative control; sh-ADAR1, shRNA specific to ADAR1; sh-ADAR2, shRNA specific to ADAR2. Values above each band represent band intensity and were calculated using ImageJ software. Band intensity in the first lane was normalized as 1.0. ND, not detected