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. 2021 Oct 29;12(3):1186–1197. doi: 10.1016/j.apsb.2021.10.023

Figure 1.

Figure 1

NF-κB activation in brown adipocytes. The primary mesenchymal cells were differentiated for 4 days in the cocktail containing 0.5 mmol/L IBMX, 125 μmol/L indomethacin, 1 μmol/L dexamethasone, 20 nmol/L insulin, and 1 nmol/L T3, and followed by additional 4 days of differentiation in medium containing 20 nmol/L insulin and 1 nmol/L T3. (A) Proteins of P65 and P50. The whole cell lysate was used in Western blot (WB) with antibodies to P65 and P50. WB signals were quantified and expressed in relative signal strength over that of Day 1. (B) mRNA levels of p65 and p50. mRNA level was determined by qRT-PCR on days of differentiation indicated. (C) NF-κB activation in BAT of WT mice after 4 h of cold challenge (n = 6). (D) Il-6 mRNA in BAT of WT mice after 4 h cold stimulation (n = 6). (E) NF-κB activation by β3 agonist. The primary brown adipocytes of WT mice were examined for NF-κB activation after a β3-adrenergic receptor agonist (CL-316243, 10 μmol/L) treatment at different times (30, 60, and 120 min) as indicated by WB. (F) Il-6 mRNA expression. mRNA level was determined by qRT-PCR. The cell culture studies were repeated three times in this study with consistent results. Representative blots are shown. Each bar represents mean ± SD (n = 6). ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001.