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Fig. 7 | BMC Biology

Fig. 7

From: Mettl14-mediated m6A modification is essential for visual function and retinal photoreceptor survival

Fig. 7

m6A depletion reduces the expression of cilium-associated genes and affects ciliogenesis in rods. A RT-qPCR verified the RNA-seq results for cilium-associated genes (n = 6). B IGV tracks displaying MeRIP-seq (lower panels, IP) and RNA-seq (upper panels, input) reads along the locus of cilium-associated genes in Ctrl and RKO retinas. Two replicates are shown. C Enrichment of m6A at multiple sites (highlighted in Fig. 7B) in transcripts of significant cilium-associated genes was determined by MeRIP-qPCR. D The left panel shows a schematic diagram of a rod photoreceptor with its ciliary region connecting the inner (IS) and outer segment (OS) compartments, which mainly consists of the connecting cilium (CC) and basal body (BB). The right panel shows representative immunofluorescence images with arrowheads pointing to the ciliary marker acetylated α-tubulin (red) and the distal appendage (DAP) marker CEP164 (green) in retinas from 3-month-old Ctrl and RKO mice. Nuclei were counterstained with DAPI (blue). Scale bars, 5 μm. Higher-magnification images of representative ciliary regions are shown on the right. Scale bars: 1 μm. E Quantitative analyses of the length of the CC in Ctrl and RKO photoreceptors (n = 10). F Quantitation of CEP164-marked DAPs in Ctrl and RKO photoreceptors (n = 9). G Representative immunoblots and quantification of acetylated α-tubulin protein levels in retinas from 3-month-old mice. Protein expression data were normalized according to GAPDH (n = 6). H Representative electron micrographs of the ciliary region of photoreceptor cells from 3-month-old Ctrl and RKO mice. Scale bars: 2 μm. Arrowheads indicated typical structure of CC. Higher-magnification images of representative ciliary regions are shown on the right. Scale bars: 500 nm. I Quantitative analyses of the length of the BB and CC in Ctrl and RKO photoreceptors (n = 7). *p < 0.05; **p < 0.01; ***p < 0.001. #, no significant difference. Data are presented as the mean ± SEM

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