Fig. 7: Conditional knockdown of METTL3 in mouse lens.
a Gene expressions in lens epithelia of Prox1-CreERTwt/wt;Mettl3flox/flox mice (WT) and Prox1-CreERTct2/wt;Mettl3flox/flox mice (cKD) (n = 9, METTL3 p < 0.0001, DVL2 p = 0.0022). b Protein expressions in lens epithelia of WT and cKD mice (n = 9, all p < 0.0001). c Immunostaining of WGA of lens fiber cell membrane in subcapsular areas at the equator of WT and cKD lens. Yellow arrows show the discontinuous cell membrane and disordered alignment. Scale bar: 10 μm. d Representative SEM images around subcapsular areas at the equator of WT and cKD lens. Left panel: The primary scam mouse lens half (F front, E equator). Red rectangles showed the ROI of formal SEM scans around subcapsular areas at the equator of lens matrix. Right panel: Representative SEM formal scans of lens fibers. e Scheme diagram depicting the timetable of tamoxifen induction and intravitreal injection of active TGF-β1. f The representative SEM scans of lens from WT, cKD, and WT or cKD mice after intravitreal injection of TGF-β1. The average lens fiber thickness measured: a random 6 × 6 μm area were chosen to measure the average lens fiber thickness in each lens scans (n = 6, WT vs. CKO p = 0.9997, WT vs. WT + TGF-β1 p = 0.0001, CKO vs. CKO + TGF-β1 p = 0.3987, WT + TGF-β1 vs. CKO + TGF-β1 p = 0.0058). Scale bar: 2 μm. g Swept-source optical coherence tomography scans of lens from WT, cKD, and WT or cKD mice after intravitreal injection of TGF-β1. Yellow rectangles showed the area of partial enlargement view. The mean intensity of the nuclear area was measured by image J. The band density in Western blot was normalized to loading control as a ratio for statistical analysis. Data present are mean ± SD. Level of significance was detected using two-sided two-way ANOVA with Tukey’s multiple comparisons test (a, b, f, g). n = biological replicates, ns. = not significant, ****p < 0.0001, ***p < 0.001, **p < 0.01 and *p < 0.05.
