Fig. 3. Phytosterol-containing PN solutions promote activation of liver macrophages and BMDMs.

(A) qRT-PCR analysis depicting relative amounts of Il6 mRNA (canonical indicator of macrophage activation) in purified hepatic CD11b⁺ cells (presented as hepatic macrophages) derived from the various groups of mice. Hepatic macrophage activation is increased in SO-PN–infused mice. DSS treatment alone with infusion of NS does not induce macrophage activation. Macrophage activation was attenuated in livers from mice infused with FO-PN and NoL-PN. Macrophage activation was also promoted in livers from mice infused with stigmasterol-spiked PN-SO solution (PN-FO-Stig3). Gene expression was determined after normalization to hypoxanthine-guanine phosphoribosyltransferase 1 (Hprt1) as an endogenous control gene. mRNA amounts are expressed relative to results obtained from untreated chow-fed control mice. Livers were pooled from three to five mice per group, and means and SEM from technical triplicates are shown. One representative of two independent CD11b⁺ cell isolations is shown. (B to D) StigAc induces transcription of the canonical macrophage proinflammatory genes Il6 (B), Il1β (C), and Tnfα (D) in naïve BMDMs. mRNA amounts are expressed relative to results obtained from untreated BMDMs. Shown are means and SEM from a technical triplicate from one representative experiment of a series of three independent experiments (BMDMs obtained from three different mice). *P < 0.05 versus all groups without *; *^#P < 0.05 versus all other groups by one-way ANOVA with Tukey correction for multiple comparisons; exact P values for each comparison are depicted in table S2.