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. 2010 Mar 9;2010:525180. doi: 10.1155/2010/525180

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Copyright © 2010 Suzanne Heemskerk et al.

This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Expression of iNOS and effect of NO on Abcb1/P-gp activity in GERP cells. ((a) n = 4) NOSII expression in GERP cells after exposure to either 10 μg/mL LPS (closed bars), 10 ng/mL TNF-α (striped bars) or both 10 μg/mL LPS and 10 ng/mL TNF-α (chequered bars) for 2, 6 or 24 hours (n = 4). mRNA levels were determined with RQ-PCR and expression was normalized for the GAPDH C_T value. ((b) n = 4) iNOS protein expression in GERP cells (control; lanes 1 and 2) after exposure to either 10 μg/mL LPS (lanes 3 and 4), 10 ng/mL TNF-α (lanes 5 and 6) or both 10 μg/mL LPS and 10 ng/mL TNF-α (lanes 7 and 8) for 6 hours. ((c) n = 3–5) P-gp transport activity in GERP cells after exposure to 10 ng/mL TNF-α or both 10 μg/mL LPS and 10 ng/mL TNF-α in combination with 50 or 10 μM of the NO-scavenger PTIO, respectively, for 24 hours compared to control (cells exposed to culture medium). ((d) n = 4) P-gp protein expression in GERP cells (control; lanes 1 and 2) after exposure to 100 μM SNP (lanes 3 and 4) for 1 hour with 5 hours recovery. ((e) n = 4) P-gp transport activity in GERP cells, compared control, after exposure to 100 or 500 μM SNP for 6 hours or for 1 hour with a subsequent recovery period of 5 hours. Total cell fractions of GERP cells were used and expression of iNOS (b) or P-gp (d) was determined by Western blotting. Relative pixel intensities (ratio iNOS/β-actin (b) and ratio P-gp/β-actin (d)) were determined through image analysis. Data are expressed as mean ± SEM. Significantly different compared to control (*: P < .05, **; P < .01, ***: P < .001).

Expression of iNOS and effect of NO on Abcb1/P-gp activity in GERP cells. ((a) n = 4) NOSII expression in GERP cells after exposure to either 10 μg/mL LPS (closed bars), 10 ng/mL TNF-α (striped bars) or both 10 μg/mL LPS and 10 ng/mL TNF-α (chequered bars) for 2, 6 or 24 hours (n = 4). mRNA levels were determined with RQ-PCR and expression was normalized for the GAPDH C_T value. ((b) n = 4) iNOS protein expression in GERP cells (control; lanes 1 and 2) after exposure to either 10 μg/mL LPS (lanes 3 and 4), 10 ng/mL TNF-α (lanes 5 and 6) or both 10 μg/mL LPS and 10 ng/mL TNF-α (lanes 7 and 8) for 6 hours. ((c) n = 3–5) P-gp transport activity in GERP cells after exposure to 10 ng/mL TNF-α or both 10 μg/mL LPS and 10 ng/mL TNF-α in combination with 50 or 10 μM of the NO-scavenger PTIO, respectively, for 24 hours compared to control (cells exposed to culture medium). ((d) n = 4) P-gp protein expression in GERP cells (control; lanes 1 and 2) after exposure to 100 μM SNP (lanes 3 and 4) for 1 hour with 5 hours recovery. ((e) n = 4) P-gp transport activity in GERP cells, compared control, after exposure to 100 or 500 μM SNP for 6 hours or for 1 hour with a subsequent recovery period of 5 hours. Total cell fractions of GERP cells were used and expression of iNOS (b) or P-gp (d) was determined by Western blotting. Relative pixel intensities (ratio iNOS/β-actin (b) and ratio P-gp/β-actin (d)) were determined through image analysis. Data are expressed as mean ± SEM. Significantly different compared to control (*: P < .05, **; P < .01, ***: P < .001).