Figure 7.

Efgartigimod does not rescue the aberrant Dsg3 staining pattern induced by hAK23 and 4B3. hTert keratinocytes were cultured on coverslips in KGM2 medium with 0.05 mM CaCl₂ for at least three days, and then switched to KGM2 with 2 mM CaCl₂ for 24 h. The cells were either mock-treated, or efgartigimod (25 µg/mL) was applied for 30 min, after which a 24 h treatment with the recombinant antibodies hAK23 (12.5 µg/mL), 4B3 (50 µg/mL), an isotype-matched human IgG4 (12.5 µg/mL) or IgG1 control (50 µg/mL) coupled to Alexa Fluor 488 (green) was initiated. After methanol fixation, the control IgG-treated cells were stained with the 5H10 anti-Dsg3 antibody, detected with a fluorochrome coupled secondary antibody (anti-mouse Alexa Fluor 546, red). Representative images from one out of at least three independent experiments are shown. Scale bar: 20 µm.