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. 2017 Sep 25;127(11):4001–4017. doi: 10.1172/JCI92893

Figure 4. mTORC1 loss of function is associated with reduced desmosomal mRNA expression and is compensated by transfection of exogenous desmosomal cadherins.

Figure 4

(A–C) Quantitative real-time reverse transcriptase PCR of desmosomal gene transcripts in P0 Rptor WT versus cKO epidermis (A); in WT versus inducible Rptor-cKO keratinocytes grown in high calcium (B); and in Rheb WT versus cKO keratinocytes grown in high calcium (C) (r = 3 for each; error bars represent SEM; P values indicated are by Student’s t test). (D) Immunoblotting of inducible Rptor-cKO keratinocytes transiently transfected with empty vector, DSG1 mouse cDNA ORF, or DSG3 mouse cDNA ORF for desmosomal proteins (DSP, DSG1, DSG3, and PKP3) and markers of biochemical differentiation (TGM1, involucrin, and loricrin). (E) Densitometry quantification of representative immunoblot experiments shown in D (r = 3; error bars represent SEM; P values by 1-way ANOVA). (F) Dispase dissociation assay for Rheb-cKO keratinocytes after transient transfection with empty vector, DSG1 mouse cDNA ORF, or DSG3 mouse cDNA ORF. Scale bar: 1 cm. (G) Quantification of representative dispase assay experiments shown in F (r = 4; error bars represent SEM; P values by 1-way ANOVA). EV, empty vector.