Journal: The Journal of cell biology
Article Title: DPM1 modulates desmosomal adhesion and epidermal differentiation through SERPINB5.
doi: 10.1083/jcb.202305006
Figure Lengend Snippet: Figure 2. DPM1 modulates DSP localization and cytoskeletal organization. (a) Immunostaining of DSP and DSG2 in HaCaT keratinocytes. Merge indicates overlap between DSP, DSG2, and nuclei stained with DAPI. Scale bar: 10 µm distance. (b) Quantification of the number of DSP puncta over the respective length of cell membrane (µm) from individual cells (represented by individual dots), N = 3. Unpaired Student’s t test. (c) Immunostaining of DSP and DSG2 in primary human keratinocytes. Scale bar: 10 µm distance. Panel shows representatives of three biological replicates. (d) Immunostaining of DSP in 3D-RHE of control (sgNT1) and sgDPM1 conditions. White dashed line indicates insert membrane. Magenta dashed rectangles mark regions magnified on the right (zoomed 2× of original image). Representative of three biological replicates. Scale bar: 10 µm distance. (e–g) Keratin staining depicted by pan-cytokeratin in
Article Snippet: The following primary antibodies were diluted with odyssey blocking buffer in trisbuffered-saline containing 0.1% Tween20 (Thermo Fisher Scientific) and incubated overnight at 4°C, with rotation: mouse Dsc2 mAb (#60239-1-Ig; Proteintech), mouse Dsg2 mAb (clone 10G11, #BM5016; Acris), rabbit Dsg3 pAb (EAP3816; Elabscience), mouse Pkp1 mAb (clone 10B2, #sc-33636; Santa Cruz), mouse Pkp2 mAb (#651101; Progen), mouse PG mAb (clone PG5.1, #61005; Progen), mouse DSP mAb (#sc-390975; Santa Cruz), mouse E-Cad mAb (clone 36, #610181; BD Biosciences), rabbit DSC1 mAb (#ab150382; Abcam), mouse DSG1 (#651111; Progen), rabbit keratin 10 (#905403; BioLegend), mouse keratin 14 (ab7800; Abcam), rabbit DPM1 (#12403-2-AP; Proteintech), rabbit SERPINB5 (MASPIN #ab182785; Abcam), rabbit mGFP (#TA150122; Thermo Fisher Scientific), mouse GAPDH mAb (clone 0411, #sc-47724; Santa Cruz), and mouse α-tubulin (clone 10D8, #627901; BioLegend).
Techniques: Immunostaining, Staining, Membrane, Control