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. Author manuscript; available in PMC: 2015 Feb 1.
Published in final edited form as: Mol Nutr Food Res. 2013 Sep 3;58(2):239–247. doi: 10.1002/mnfr.201300181

Figure 2. Rosiglitazone regulates NF-κB activity in Nfkb-RE/GFP adipocytes in LPS-dependent manner.

Figure 2

(a) Nfkb-RE/GFP preadipocytes (ND, white bar) were differentiated for 4d (D, black bar) and stimulated with and without 100nM BRL for 24h (hatched bar). The GFP/mCherry ratio was obtained from total cell lysates in RIPA buffer (Mann Whitney U test, n=3 or 4, mean±SD).

(b) Relative activation of NF-κB in Nfkb-RE/GFP preadipocytes differentiated for 4 days and stimulated with different concentration of BRL. Data are shown as relative activation of NF-κB in Nfkb-RE/GFP preadipocytes (ND, GFP/mCherry ratio 100%) compared to adipocytes treated with BRL (Mann Whitney U test, n=4 or 8, mean±SD)

(c) Relative activation of NF-κB in Nfkb-RE/GFP preadipocytes differentiated for 4d and simultaneously stimulated with different concentration of BRL and 10ng/mL LPS for 24h (dashed line). Data are shown as GFP/mCherry ratios. An asterisk (P<0.01), signifies a significant difference between differentiated Nfkb-RE/GFP stimulated with BRL and LPS vs. LPS-stimulated control (Mann Whitney U test, n=4, mean±SD).

(d) MCP-1 levels in adipocyte treated with and without LPS. Experiments were performed as in b and c. Differentiated for 3d adipocytes were pre-treated for 10 min with BRL, then LPS (10 ng/mL) was added for 24 h. Stimulation media contained 1%FBS. Media and cell lysates were collected for the measurement of MCP-1 protein level by ELISA and Nfkb-RE/GFP/mCherry fluorescence ratio, respectively. Insert shows the correlation between MCP-1 protein levels and Nfkb-RE/GFP fluorescence normalized by mCherry. Pearson correlation test.