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. 2018 Mar 9;9(3):389. doi: 10.1038/s41419-018-0380-9

Fig. 1. TNF-α and IFN-γ promote Dsg2 intracellular and extracellular cleavage.

Fig. 1

a Model of Dsg2 with mapped epitopes for AH12.2 (extracellular [N-term] specific) and 4B2 (intracellular [C-term] specific) monoclonal antibodies. EC extracellular cadherin domain, EA extracellular anchor, TM transmembrane domain, IA intracellular anchor, ICS intracellular cadherin-typical sequence, IPL intracellular proline-rich linker domain, RUD repeated unit domain, DTD desmoglein-specific terminal domain. b Top. Western blot using an antibody against Dsg2 C-term of cell lysates of T-84s treated with TNF-α and IFN-γ for 24 h. Bottom. Densitometry of blots. Density for all bands in each lane was collected. The ratio of the density of the ~55 kDa Dsg2 ICF band to the density of all bands for a given lane was then calculated. The data in the graph is the average of these ratios within each experimental group. *p = 0.0067; #p = 0.0003, n = 11 per group. For full lanes of Dsg2 C-term blot, please see Supplemental Fig. 2A. c Western blot using an antibody against Dsg2 N-term of cell culture supernatants of T-84s treated as in Fig. 1b. For full lanes of Dsg2 N-term blot please, see Supplemental Fig. 2B. d Top. Western blot using an antibody against Dsg2 C-term of cell lysates of T-84s treated with TNF-α and IFN-γ in the presence or absence of GI254023X for 24 h. Bottom. Densitometry of blots. Densitometry collected and analyzed as for Fig. 1b. n ≥ 4 per group. e Western blot using an antibody against Dsg2 N-term of cell culture supernatants of T-84s treated as in Fig. 1d. f Western blot using an antibody against Dsg2 C-term of cell lysates of T-84s treated with TNF-α and IFN-γ for 6, 12, or 24 h. g Western blot using an antibody against Dsg2 N-term of T-84 cell culture supernatants of cells treated as in Fig. 1f. All blots are representative of at least three independent experiments