Figure 1

Figure 1 Identification of FTO as a potential regulator of DR

(A) m⁶A dot blot assay of global m⁶A abundance in HUVECs treated with normal glucose (5 mM, NG) or high glucose (25 mM, HG) using 800 or 400 ng total RNAs. MB staining is used as a loading control. n = 3 per group. (B) m⁶A RNA methylation quantification assay of global m⁶A abundance in HUVECs treated with NG or HG. n = 3 per group. (C) m⁶A dot blot assay of global m⁶A abundance in retina originated from control or STZ mice using 800 or 400 ng total RNAs. MB staining is applied as a loading control. n = 3 per group. (D) m⁶A RNA methylation quantification assay of global m⁶A abundance in retina originated from control or STZ mice. n = 3 per group. (E) qPCR shows mRNA levels of m⁶A regulators METTL3, METTL14, WTAP, ALKBH5 and FTO in HUVECs treated with NG or HG. n = 3 per group. (F) qPCR presents mRNA levels of m⁶A regulators Mettl3, Mettl14, Wtap, Alkbh5 and Fto in retina originated from control or STZ mice. n = 3 per group. (G) Immunoblotting of METTL3, ALKBH5 and FTO in HUVECs treated with NG or HG. GAPDH is used as an internal control. n = 3 per group. (H) Immunoblotting of Mettl3, Alkbh5 and Fto in retina originated from control or STZ mice. GAPDH is used as an internal control. n = 3 per group. (I) m⁶A dot blot assay of global m⁶A abundance in HUVECs transduced with L-EV or L-FTO using 800 or 400 ng total RNAs. MB staining is applied as a loading control. n = 3 per group. (J) Immunofluorescence staining of FTO in FVMs obtained from PDR patients or ERMs isolated from age matched controls without diabetes. Fundus photographs and OCT images of PDR patients or age matched controls with ERM are shown. n = 35, ERM; n = 33, PDR. Scale bar: 50 µm. Data information: Data represent different numbers (n) of biological replicates. Data are shown as mean ± SEM. Two-tailed Student’s t test is used. NS: not significant (p > 0.05); *p < 0.05; **p < 0.01; and ***p < 0.001. Source data are available online for this figure.