Fig. 8: Tmem67ΔCLE/ΔCLE MEFs respond to canonical and non-canonical Wnt signaling. | Nature Communications

Fig. 8: Tmem67ΔCLE/ΔCLE MEFs respond to canonical and non-canonical Wnt signaling.

From: Cleavage of the Meckel-Gruber syndrome protein TMEM67 by ADAMTS9 uncouples Wnt signaling and ciliogenesis

Fig. 8

a–d Tmem67ΔCLE/+ and Tmem67ΔCLE/ΔCLE MEFs stained for active β-catenin (green) and F-actin (Phalloidin, red) upon L-cell (control), Wnt3a, and Wnt5a conditioned medium treatment. Mean nuclear active β-catenin signal from 20 nuclei from each group is shown (n = 3 independent experiments), **** indicates a p-value <0.0001, ***<0.001 (p = 0.0002) in one-way Kruskal–Wallis test with a Dunn’s multiple comparison test in (c). Mean F-actin pixel intensity from 4 different fields of cells in each group is shown in (d) (n = 4 independent experiments), *** indicates a p-value <0.001(p = 0.0002), **<0.01(p = 0.0021), *<0.05(p = 0.0158) in a one-way ANOVA with Tukey’s multiple comparisons test. Error bars indicate Mean ± S.D. in (c and d). e qRT-PCR for indicated genes in Wt, Tmem67ΔCLE/ΔCLE, and Tmem67-null MEFs treated with L-cell (blue), Wnt3A (purple), or Wnt5a (green) conditioned medium. **** indicates a p-value <0.0001, ***<0.001, **<0.01, *<0.05 in two-tailed unpaired t-test. Error bars indicate Mean ± S.E.M. Data represent 3 independent experiments. p = 0.0002(Jun, Wt+Wnt3a), p = 0.1135(Myc, Wt+Wnt3a), p = 0.8644(Ccnd1, Wt+Wnt3a), p = 0.0001(Lef1, ΔCLE+Wnt3a), p = 0.0099(Jun, ΔCLE+Wnt3a), p = 0.0002(Myc, ΔCLE+Wnt3a), p = 0.6816(Ccnd1, ΔCLE+Wnt3a), p = 0.0045(Tcf7, KO+Wnt3a), p = 0.0099(Lef1, KO+Wnt3a), p = 0.5507(Myc, KO+Wnt3a), p = 0.0241(Jun, ΔCLE+Wnt5a), p = 0.0009(Axin2, ΔCLE+Wnt5a), p = 0.0323(Tcf7, KO+Wnt5a), p = 0.0101(Jun, KO+Wnt5a), p = 0.0013(Myc, KO+Wnt5a), p = 0.0016(Ccnd1, KO+Wnt5a). f qRT-PCR for indicated genes in Wt, Tmem67ΔCLE/ΔCLE and Tmem67-null MEFs treated with L-cell (blue) or Wnt5A (green) medium * indicates a p-value <0.05 in two-tailed unpaired t-test. Error bars indicate Mean ± S.E.M. Data represent 3 independent experiments. p = 0.0337(Plod2, Wt+Wnt5a), p = 0.0311(Hadh, Wt+Wnt5a), p = 0.0328(Plod2, ΔCLE+Wnt5a), p = 0.0321(Lcor, ΔCLE+Wnt5a). g qRT-PCR for Tmem67 in Wt and Tmem67ΔCLE/ΔCLE MEFs treated with Wnt5A (green) or Wnt3A (purple). *** indicates a p-value <0.001, *<0.05 in in two-tailed unpaired t-test. Error bars indicate Mean ± S.E.M. Data represent 3 independent experiments. p = 0.0262(Wt+Wnt3a), p = 0.0227(ΔCLE+Wnt3a), p = 0.0003 (ΔCLE+Wnt5a). Scale bars in (a) are 50 μm and 5 μm.

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