Loss of DPM1 leads to reduced oligomannose structures in keratinocytes. (a) Western blot showing DPM1 in sgDPM1, sgDPM2, and sgDPM3 HaCaT keratinocytes. α-Tubulin was used as internal loading control. (b) Mass-spectrometry analysis of N-glycome from sgNT1 and sgDPM1 cell lysates. X-axis denotes mass to charge (m/z) ratio and y-axis denotes the intensities of the corresponding m/z peaks (A.U.). The m/z peaks denote the respective glycan structures as depicted and the monosaccharide codes are provided in the color-coded legends. Cyan box marks oligomannose structures in sgNT1 conditions that are drastically reduced in sgDPM1. (c and d) Western blot images and quantifications of desmosomal proteins from sgDPM1, sgDPM2, and sgDPM3 HaCaT keratinocytes. Representative images of three biological replicates are shown. GAPDH was used as internal loading control (N = 3). (e) Immunofluorescence staining of DPM1 and filaggrin in 3D-RHE and human foreskin tissue, as indicated. Filaggrin was used as a differentiation marker. Dashed line indicates insert membrane/basement membrane. Scale bar: 10 µm. Panel shows representative of three biological replicates. (f) Immunostaining of CK10 and filaggrin in 3D-RHE and human foreskin tissue, as indicated. Dashed line indicates insert and basement membrane. Scale bar: 10 µm distance. Panel shows representative of three biological replicates. Source data are available for this figure: SourceData FS1.