Figure S9. Homologies of the identified Msi2 binding motif in mouse 3’-UTR with Shh mRNA from other species. (a) Distance tree view of the identified Msi2 binding motif Shh in 3’-UTR across species. Unknown indicates Msi2 binding motif sequence of mouse Shh (b) List of homologies of the Msi2 binding motif in 3’-UTR by BLASTS pairwise Shh. Shh alignment. Figure S10. Msi2 represses the Hh signaling. (a) hybridization for and IHC for In situ Shh Msi2 in control and DTG HFs 36 and 3 days post-depilation. (b) hybridization for In -situ and IHC for Msi2 in control and cKO HFs 36 hours and 3 days post-depilation. (c) IHC Shh for Gli1 in control and DTG, or in control and cKO HFs at P24 3 days post-depilation. Control and DTG mice were treated with Dox at P18 and depilated at P21. Control and cKO mice were depilated at P21. Scale bar: 50 m. (d) Western blotting for Gli1 in control and DTG skin 36 hours post-depilation at P21. Control and DTG mice were treated with Dox from P18 and depilated at P21. Control mice were topically treated with a single 25 L application of vehicle (95% acetone/5% DMSO), while DTG mice were given a single 25 L topical application of SAG (0.06 g per L in vehicle, 120 mM). Second application was done 24 hours later. Ve: vehicle. SUPPLEMENTARY MATERIALS AND METHODS Mouse strains and mice were previously described (; ). and TRE-MSI2 Msi2flox/flox K14-rtTA mice were obtained from Jackson Laboratory (stock numbers 007678 and 004782).
Figure S9. Homologies of the identified Msi2 binding motif in mouse Shh 3’-UTR with Shh mRNA from other species. (a) Distance tree view of the identified Msi2 binding motif in Shh 3’-UTR across species. Unknown indicates Msi2 binding motif sequence of mouse Shh. (b) List of homologies of the Msi2 binding motif in Shh 3’-UTR by BLASTS pairwise alignment. Figure S10. Msi2 represses the Hh signaling. (a) In situ hybridization for Shh and IHC for Msi2 in control and DTG HFs 36 and 3 days post-depilation. (b) In -situ hybridization for Shh and IHC for Msi2 in control and cKO HFs 36 hours and 3 days post-depilation. (c) IHC for Gli1 in control and DTG, or in control and cKO HFs at P24 3 days post-depilation. Control and DTG mice were treated with Dox at P18 and depilated at P21. Control and cKO mice were depilated at P21. Scale bar: 50 m. (d) Western blotting for Gli1 in control and DTG skin 36 hours post-depilation at P21. Control and DTG mice were treated with Dox from P18 and depilated at P21. Control mice were topically treated with a single 25 L application of vehicle (95% acetone/5% DMSO), while DTG mice were given a single 25 L topical application of SAG (0.06 g per L in vehicle, 120 mM). Second application was done 24 hours later. Ve: vehicle. SUPPLEMENTARY MATERIALS AND METHODS Mouse strains TRE-MSI2 and Msi2flox/flox mice were previously described (; ). K14-rtTA and mice were obtained from Jackson Laboratory (stock numbers 007678 and 004782).
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