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. 2018 Feb 8;293(12):4582–4590. doi: 10.1074/jbc.RA117.001343

Figure 2.

Figure 2.

MCPyV ST up-regulates CLIC proteins. A, quantitative proteomic analysis shows an increase in CLIC family protein levels upon MCPyV ST expression. B, i293-EGFP or i293-EGFPST cells were incubated for 48 h in the presence of doxycycline, whereas MCC13 cells were transfected with EGFP or EGFP-ST for 24 h. Cell lysates were probed using CLIC1- and CLIC4-specific antibodies. GAPDH and β actin were used to ensure equal loading. The 2T2 hybridoma was used to probe for MCPyV ST expression. C, densitometry was performed using ImageJ (n = 3) for (i) i293 or (ii) MCC13 samples. *, p < 0.01. D, total RNA was extracted from induced i293-EGFP or i293-EGFP-ST cells, reverse transcribed, and RT-quantitative PCR performed. Transcript levels were analyzed using the comparative CT method (n = 3). *, p < 0.01. E, (i) surface biotinylation experiments were performed in induced i293-EGFP or i293-EGFP-ST cells; lysates were probed for CLIC1, CLIC4, and CD71 as a surface marker control. (ii) densitometry of Western blots was performed using ImageJ software (n = 3). *, p < 0.01.