Fig. 8
From: Malate initiates a proton-sensing pathway essential for pH regulation of inflammation
BiP-IRF2BP2 signaling links pH fluctuation with inflammation. (a, c) Il1b mRNA levels (a) and pro-IL-1β protein abundance (c) in BMDMs isolated from Hspa5f/f or Hspa5f/f; Lyz2-Cre mice treated with or without acidized medium (AcM) (pH 6.7), under stimulation of LPS for 24 h. (b) Il1b mRNA levels by qPCR in BMDMs isolated from Irf2bp2f/f or Irf2bp2f/f; Lyz2-Cre mice treated with or without acidized medium (AcM) (pH 6.7), under stimulation of LPS for 24 h. (d, f) Protein abundance of pro-IL-1β (d) and mRNA levels of Il1b (f) in BMDMs isolated from Irf2bp2f/f or Irf2bp2f/f; Lyz2-Cre mice treated with or without Monensin (1 μM), under stimulation of LPS for 24 h. (e) Il1b expressions in BMDMs isolated from Hspa5f/f or Hspa5f/f; Lyz2-Cre mice treated with or without monensin (1 μM), under stimulation of LPS for 24 h. (g, h) pro-IL-1β protein levels in BMDMs isolated from Hspa5f/f or Hspa5f/f; Lyz2-Cre mice treated with or without NH4Cl (15 mM), under stimulation of LPS for 16 h. (i) Il1b mRNA levels in BMDMs isolated from Hspa5f/f or Hspa5f/f; Lyz2-Cre mice treated with or without NH4Cl (15 mM), under stimulation of LPS for 8 h. (j) Il1b mRNA levels in BMDMs isolated from Irf2bp2f/f or Irf2bp2f/f; Lyz2-Cre mice treated with or without NH4Cl (15 mM), under stimulation of LPS for 8 h. (k, l) pro-IL-1β protein levels in BMDMs isolated from Irf2bp2f/f or Irf2bp2f/f; Lyz2-Cre mice treated with or without NH4Cl (15 mM), under stimulation of LPS for 16 h. (m) Representative H&E-stained sections of colons from Hspa5f/f, Hspa5f/f; Lyz2-Cre mice sampled on day 6 after the saline or NaOH exposure. Scale bar 100 μm. (n) Length of colons from Hspa5f/f or Hspa5f/f; Lyz2-Cre mice at day 6 of NaOH-induced colitis model (n = 7-9 per group). (o, p) mRNA levels of Il1b (o) and Il6 (p) in colons collected at day 3 after the saline or NaOH exposure, analyzed by qPCR. (q) Immunoreactive staining (brown) for IRF2BP2 [scale bar, 100 (left) or 50 (right) μm]. (r) Western blot of IRF2BP2 in colons of mixed sample from Hspa5f/f or Hspa5f/f; Lyz2-Cre mice at day 3 of NaOH-induced colitis model (n = 8 per group). The “X” means unknown proteins detected in immunoblotting visualizing IRF2BP2. n ≥ 3. Data are shown as mean ± SEM. *p < 0.05; **p < 0.01; ***p < 0.001 (unpaired Student’s t-test)
