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Figure 3 PPAR Is Required for Pioglitazone-Mediated Repression of TNF-Induced Endothelial VCAM-1 mRNA Expression

Northern blot analysis was performed on total RNA isolated from PPAR^–/– endothelial cells transfected either with PPAR-containing pSG5 overexpression vector (A) or pSG5 alone (B) before stimulation with TNF in either the absence or presence of WY (100 µM) or pioglitazone (10 µM), with subsequent probing for VCAM-1 or GAPDH expression. (C) Quantification of the VCAM-1 mRNA response to pioglitazone relative to GAPDH mRNA expression levels (n = 3, ^#p < 0.05 TNF vs. vehicle; *p < 0.05 pioglitazone/TNF vs. TNF alone; p < 0.05 WY14643/TNF vs. TNF alone, Mann-Whitney U test). (D) Cells were transfected in panels A and B but with a concentration gradient of pSG5-PPAR as shown before TNF stimulation in either the absence or presence of pioglitazone 10 µM (n = 3, *p < 0.05 pioglitazone/TNF vs. TNF alone). Abbreviations as in Figure 1.