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. 2020 May 1;15(5):e0232630. doi: 10.1371/journal.pone.0232630

Fig 7. Effect of BBR on PA and LPS-induced activation of inflammation, UPR, and ERK in primary mouse hepatocytes.

Fig 7

Primary mouse hepatocytes were isolated from C57BL/6 wild type mice and pre-treated with BBR (5 μM) for 1 h, then treated with PA (0.25mM) and LPS (25 ng/mL) for 6 h. At the end of treatment, cell culture medium and total cellular protein were collected. The protein levels of TNF-α, IL-6, and MCP-1 were determined by ELISA and were normalized by total protein amounts and expressed as pg/mg of protein. Total cell lysates were prepared for western blot analysis for IL-1β, CHOP, ATF4, XBP-1, phospho(p)-ERK, total (T)-ERK, and β-Actin. Values are mean ± S.E. of three independent experiments. Statistical significance relative to vehicle control, *p<0.05, **p<0.01; relative to PA+LPS, #p<0.05. A. The relative protein levels of TNF-α, IL-6, and MCP-1; B. Representative immunoblots of IL-1β and β-Actin; C. The relative protein levels of IL-1β; D. Representative immunoblots of CHOP, ATF4, XBP-1s, and β-Actin. E. The relative protein levels of CHOP, ATF4, and XBP-1s; F. Representative immunoblots of phospho(p)-ERK and total (T)-ERK. G. The relative protein levels of p-ERK/total ERK.