Fig. 3

CD98hc is required for proper skin mechanical homeostasis in vivo. a Representative skin sagittal sections of CD98hcfl/fl or Fsp1Cre, CD98hcfl/fl mice stained with H&E. Scale bar is 50 µm. b Genetic deletion of CD98hc in dermal fibroblasts alters dermal mechanical properties. Young’s elastic modulus (E) of the dermis was measured by AFM with a spherical probe (mean with s.d. as error bars, n = 5 and 4 mice, respectively, ^*P < 0.05 in a Student’s t test). c E magnitude heatmaps of areas of dermis scanned by AFM using a pyramidal probe. d, e Loss of CD98hc in dermal fibroblasts reduces collagen assembly as observed by picrosirius red staining under transmitted or polarized light (d), fiber length (e) (mean with s.d. as error bars, at least 300 fibers were counted with n = 3 ^**P < 0.01 in a Welch’s t test (e)). f Relative mRNA levels of LOX was measured by RT-qPCR on total mRNA extracted from skin of CD98hcfl/fl or Fsp1Cre, CD98hcfl/fl mice (mean with s.d. as error bars, n = 6 ^****P < 0.0001 in a Student’s t test). g, h Loss of CD98hc in dermal fibroblasts impairs elastin fibers assembly as observed on skin sagittal sections stained for elastin. Scale bar is 50 µm. Means are plotted with s.d. as error bars. At least 300 fibers were measured on sections from n = 3 mice. ^****P < 0.0001 in a Student’s t test. i Relative mRNA levels of LOX was measured by RT-qPCR on total mRNA extracted from control or CD98hc-null dermal fibroblasts grown FN-coated hydrogels of indicated stiffness for 24 h. n = 2, means are plotted with s.e.m. as error bars, ^**P < 0.01, ^****P < 0.0001 in a Student’s t test. j Total cell lysates from control or CD98hc-null dermal fibroblasts grown FN-coated hydrogels of indicated stiffness for 24 h were resolved by SDS–PAGE and analyzed by Western blotting. k Relative LOX activity was measured in cell lysates from control or CD98hc-null dermal fibroblasts grown FN-coated hydrogels of indicated stiffness for 24 h. n = 2, means are plotted with s.e.m. as error bars. ^**P < 0.01, ^***P < 0.001 in a two-way ANOVA