Extended Data Fig. 7: Reduced formation of 5hmC causes acquisition of a synthetic phenotype in lung SMCs.

a–c, EM images of bronchial SMCs, 6 months after tamoxifen injection (a) and of lung vascular SMCs 8 weeks (b) or 6 months (c) after tamoxifen injection in control and Tet3^smKO mice (n = 4). Arrows point to dense bodies (control) and rough ER (Tet3^smKO). Scale bar: 1000 nm. d, Microarray expression analysis of laser capture micro-dissected bronchiolar (left panel) or vasculature (middle and right panels) regions from control and Tet3^smKO lungs, 8 weeks after tamoxifen injection (n = 3; two-sided moderated t-test with Benjamini-Hochberg correction for multiple test). Fold change refer to the Tet3^smKO/control. e, RT-qPCR analysis of miR-145 expression in SMCs from control and Tet3^smKO:T lungs (n = 4 independent animals; two-tailed unpaired t-test: *P = 0.0235). f, Quantification of protein levels of α-SMA (n = 5), MYH11 (n = 3), TAGLN (n = 3), VIM (control n = 4, Tet3^smKO:T n = 3), TPM4 (n = 3) in SMCs from control and Tet3^smKO:T lungs (two-tailed unpaired t-test: **P = 0.002, **P= 0.0045, **P = 0.0097, *P = 0.0283, *P = 0.0426). g, Immunofluorescence staining for α-SMA and Ki67 on cryosections from control and Tet3^smKO lungs, 8 weeks after tamoxifen injection (n = 3). DNA was stained by DAPI. Scale bar: 50 μm. h, Senescence-associated β-galactosidase staining of cryosections from control and Tet3^smKO lungs, 8 weeks after tamoxifen injection (n = 3). Scale bar: 50 μm. i, RT-qPCR analysis of Cdkn1a (p21), Cdkn2a (p16) in SMCs isolated from control and Tet3^smKO:T lungs 8 weeks after tamoxifen injection (n = 3 independent animals; two-tailed unpaired t-test: **P = 0.008, *P = 0.0246). Data in (e, f, i) are presented as mean values ± s.e.m.

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