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. 2020 Aug 26;9:e56980. doi: 10.7554/eLife.56980

Figure 3. Loss of m6A results in diminished WNT signaling and signs of perturbed HF fate.

(A) Left panel: representative images from E17.5 sagittal sections immunolabeled for LEF1 and PCAD (scale bars: 50 µm). White solid lines denote skin surface and dashed lines denote dermal-epidermal border. Right panel: quantification of LEF1 immunofluorescence at indicated compartments (for each condition n = 3 biological replicates ×7 images per replicate, *p<0.05 and ***p<0.001 by unpaired two-tailed Student’s t-test). (B) Left panel: representative images from P0 sagittal sections immunolabeled for LEF1 and PCAD (scale bars: 50 µm). Solid and dashed lines as in (A). Right panel: quantifications of LEF1 immunofluorescence in indicated compartments (for each condition n = 3 biological replicates ×7 images per replicate, *p<0.05 and ***p<0.001 by unpaired two-tailed Student’s t-test). (C) Representative images from P0 sagittal sections with immunohistochemistry staining of β-catenin, counter-stained with hematoxylin (scale bars: 50 µm). (D) Representative images from P0 sagittal sections stained with X-gal and nuclear fast red to examine the expression of the Gli1-LacZ transgene, a proxy for SHH signaling (scale bars: 100 µm). (E) Representative images from P6 sagittal sections stained with Oil Red O and counterstained with hematoxylin to visualize signs of HF to sebocyte fate switching within the epithelium (scale bars: 100 µm). The staining in the control dermis reflects adipocyte-derived lipids, missing in the cKO pups, which lose weight after birth. (F) Representative pictures of control (Ctrl) and cKO back skins engrafted onto Nude (Nu/Nu) mice and analyzed 15 days later. (G) Representative Hematoxylin and eosin (H and E) stained sagittal sections of 15-dayengrafted back skins (scale bars: 100 µm). (H) Representative sagittal sections as in (G) and counterstained with Oil Red O and hematoxylin (scale bars: 100 µm).

Figure 3—source data 1. Quantification of LEF1 immunofluorescence signals at E17.5 in (A).
Figure 3—source data 2. Quantification of LEF1 immunofluorescence signals at P0 in (B).

Figure 3.

Figure 3—figure supplement 1. Examination of METTL3 and YFP expression in the grafted skin.

Figure 3—figure supplement 1.

Images of back skin engrafted to Nude (Nu/Nu) mice for 15 days immunolabeled for PCAD, METTL3 and YFP demonstrating in the grafted tissue METTL3 is still largely depleted in the cKO epithelium (scale bars: 100 µm). White solid lines indicate skin surface and dashed lines indicate epithelial-dermal border.