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Figure Lengend Snippet: Relief of macrophage-mediated inflammation during AP by B. animalis colonization or lactate administration requires TLR4-MyD88 signaling. (a) Representative western blot showing expression of NF-κB signaling pathway-related molecules in the pancreas and spleen of mice treated with PBS, Abx, or Abx-treated mice colonized by B. animalis or given lactate gavage at 12 h post-induction of SAP (All experiments were repeated independently three times. one representative experiment was shown.) (b) Serum amylase activity, (c, d) pancreatic tissue lesions and (g, h) pathology scores of Tlr4 −/− or Myd88 −/− mice or their respective wild-type (WT) parent strain, treated with PBS, Abx or Abx-treated mice colonized by B. animalis or given lactate gavage at 12 h post-induction of SAP (n = 5). Expression (ELISA) of IL-1β, IL-6 and TNF-α in peritoneal macrophages extracted from (e) Tlr4 −/− or (f) Myd88 −/− mice or their respective WT parents. Cells were pretreated or not with lactic acid (15 mM), and subsequently stimulated with LPS (200 ng/mL). Data are from two independent experiments, and P values were determined by unpaired two-tailed Student’s t-test. *: P < .05; **: P < .01; ***: P < .001; ****: P < .0001.
Article Snippet: The following antibodies were used for immunoblotting, immunofluorescence, and flow cytometry analysis: anti-Caspase-1 (ab179515; Abcam), anti-NLRP3 (ab263899; Abcam), anti-ASC (sc514414; Santa Cruz), anti-NF-κB P50 (ab32360; Abcam), anti-NF-κB P65 (cst6956; Cell Signaling Technology), anti-phospho–NF-κB P65 (cst3033; Cell Signaling Technology),
Techniques: Western Blot, Expressing, Activity Assay, Enzyme-linked Immunosorbent Assay, Two Tailed Test