Skip to main content
. 2016 Jul 4;6:29253. doi: 10.1038/srep29253

Figure 5. VPA and TSA do not increase the expression of Pgp or Pgp-EGFP.

Figure 5

Expression of endogenous Pgp in wt-hCMEC/D3 cells or hCMEC/D3-MDR1-EGFP cells (in the absence of doxycycline) was not modified by treatment with the HDAC inhibitors VPA and TSA. HCMEC/D3 cells (a) or hCMEC/D3-MDR1-EGFP cells (b) were treated with VPA (300 μM) or the HDAC inhibitor TSA (330 nM) for 24 hours. Detergent extracts were analyzed by Western blotting using anti-GFP antibodies to detect GFP-tagged Pgp and anti-actin. Pgp and actin bands of three independent experiments were analyzed densitometrically and Pgp signals were normalized versus actin. It should be noted that different gels were used for the experiments with hCMEC/D3 and hCMEC/D3-MDR1-EGFP cells, so that small differences in protein migration may be missed. When both cell types were run on the same gel, hCMEC/D3 wild type cells expressed a 170-kDa protein band corresponding to endogenous Pgp whereas non-induced hCMEC/D3-MDR1-EGFP cells expressed an approximately 200-kDa protein band corresponding to the predicted size of the chimeric MDR1-EGFP21.