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. 2022 Jul 21;13:905211. doi: 10.3389/fimmu.2022.905211

Figure 2.

Figure 2

Phenotype of hRPE cells after silencing METTL3. (A) Transfection efficiency of the METTL3-silenced lentivirus in hRPE cells (BF, bright field; EGFP, enhanced green fluorescence protein; scale bar, 10 μm). (B) Silencing efficiency of METTL3 in hRPE cells. Left, representative western blotting images of METTL3. shMET, shMETTL3; shNC, Negative Control. Right, quantification of relative expression of METTL3 (mean ± SD; **** P<0.0001; n=3/group; unpaired Student’s t-test). (C) m6A% after silencing METTL3 in hRPE cells (mean ± SD; ** P<0.01; n=3/group; unpaired Student’s t-test). (D) Cell viability of hRPE cells after silencing METTL3 with or without LPS stimulation (mean ± SD; * P<0.05, *** P<0.001, **** P<0.0001; n=6/group; unpaired Student’s t-test). (E, F) Proliferation of hRPE cells. (E) Representative Edu images of hRPE cells after silencing METTL3 with or without LPS stimulation (scale bar, 166.4 μm). (F) Quantification of Edu positive cells (mean ± SD; * P<0.05, *** P<0.001; n=3/group; one-way ANOVA). (G) Protein levels of Occludin and ZO-1 in hRPE cells after silencing METTL3 with or without LPS stimulation. Upper, representative western blotting images of Occludin and ZO-1. Lower, quantification of relative expression of Occludin and ZO-1 (mean ± SD; ns P>0.05, * P<0.05, ** P<0.01, **** P<0.0001; n=3/group; unpaired Student’s t-test). (H–J) The secretion levels of IL-1β, IL-6 and IL-8 in hRPE cells after silencing METTL3 with or without 24 hours of LPS stimulation (mean ± SD; * P<0.05, ** P<0.01, *** P<0.001, **** P<0.0001; n=4-6/group; one-way ANOVA).