ADAM 10 and 17 localisation at the cell surface is increased upon MCPyV ST expression.

(A) HEK 293 cells were transfected with 1 μg of EGFP or EGFP-ST expression plasmids. 24 hours later cells were fixed and GFP fluorescence analysed by direct visualisation, whereas endogenous (i) ADAM 10 and (ii) ADAM 17 were identified by indirect immunofluorescence using specific antibodies. (B) Surface biotinylation experiments were performed in EGFP or EGFP-ST transfected HEK 293 cells, lysates were probed for ADAM 10, ADAM 17 and CD71 as a surface marker control. (C) Densitometry of immunoblots was performed using ImageJ software. Data analysed using three replicates per experiment, n = 3 and statistical analysis using a two-tailed t-test with unequal variance, *** = p<0.001, ** = p<0.01. (D) EGFP or EGFP-ST transfected HEK 293 cells were harvested and stained with (i) ADAM 10-, (ii) ADAM 17- and CD71-specific antibodies. Mean fluorescence intensity was analyzed using FlowJo software. (E) Fold difference of cell surface staining was calculated using three replicates per experiment, n = 3 by a two-tailed t-test with unequal variance, *** = p<0.001 and ** = p<0.01.