MCPyV ST expression increases the levels of ADAM proteins.

(A) i293-ST cells remained uninduced or were incubated for 48 hours in the presence of doxycycline hyclate or (C) MCC13 cells were transfected with 1μg of pEGFP or pEGFP-ST for 12 hours. Cell lysates were then probed with ADAM 10-, ADAM 17- and ADAM TS1-specific antibodies. GAPDH was used as a measure of equal loading, the 2T2 hybridoma was used to confirm MCPyV ST expression. (B and D) Densitometry quantification of immunoblots was carried out using the Image J software and is shown as a percentage relative to the loading control, GAPDH. 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<0.0001. (E) Total RNA was extracted from EGFP or EGFP-ST transfected (i) HEK 293 and (ii) MCC13 cells and relative transcript levels were analysed by qRT-PCR using GAPDH as a reference. Fold increase was determined by ΔΔCt and statistical significance analysed using a non-paired t-test, *** = p<0.001, ** = p<0.01.