Journal: Physiological Reports

Article Title: Interleukin‐38 suppresses abdominal aortic aneurysm formation in mice by regulating macrophages in an IL1RL2 ‐p38 pathway‐dependent manner

doi: 10.14814/phy2.15581

Figure Lengend Snippet: The mechanism how interleukin‐38 suppresses macrophage activation. (a) Representative western blot analysis of p‐p38, p38, p‐JNK, and JNK in the aortic wall on day 14. (b) Bar graphs showing the relative densitometry of p‐p38 ( n = 6, p = 0.0280 [ t test], p = 0.0411 [Mann–Whitney U test]) and p‐JNK ( n = 6, p = 0.6893 [ t test], p = 0.6991 [Mann–Whitney U test]). (c) Representative western blot analysis of p‐p38, p38, p‐JNK, and JNK in macrophages after TNF‐α stimulation with or without IL‐38. (d) Bar graphs showing the relative densitometry of p‐p38 ( n = 6, p < 0.0001 [ANOVA], p < 0.0001 [post hoc, Control vs. TNF‐α], p = 0.0234 [post hoc, Control vs. TNF‐α + IL‐38], p = 0.0125 [post hoc, TNF‐α vs. TNF‐α + IL‐38]) and p‐JNK ( n = 6, p = 0.0283 [ANOVA], p = 0.0264 [post hoc, Control vs. TNF‐α]). (e) Representative western blot analysis of iNOS, MMP‐9, MMP‐2, and GAPDH in macrophages after TNF‐α stimulation with or without IL‐38 in the presence of SB203580. (f) Bar graphs showing the relative densitometry of iNOS ( n = 3, p = 0.2000 [Mann–Whitney U test]), MMP‐9 ( n = 3, p = 0.7000 [Mann–Whitney U test]), and MMP‐2 ( n = 3, p = 0.7000 [Mann–Whitney U test]). (g) Representative photomicrographs of the aortic wall on day 14 in the presence of SB203580, stained with EVG (upper row) or anti‐CD68 antibody (lower row). (h) Bar graph showing the number of CD68 + cells in the aortic wall on day 14 in the presence of SB203580 ( n = 5–6, p = 0.6623 [Mann–Whitney U test]). (i) Bar graphs showing the aorta diameter ( n = 5–6, p = 0.2468 [Mann–Whitney U test]), AAA incidence ( n = 5–6, p = 0.2506 [χ 2 test]), adventitia ( n = 5–6, p > 0.9999 [Mann–Whitney U test]), intramedia ( n = 5–6, p = 0.1710 [Mann–Whitney U test]), and elastin degradation score ( n = 5–6, p = 0.4917 [ t test], p = 0.6753 [Mann–Whitney U test]) on day 14 in the presence of SB203580.

Article Snippet: Primary antibodies included anti‐TNF‐α (rabbit monoclonal, dilution 1:400, #11948; Cell Signaling Technology), anti‐iNOS (rabbit monoclonal, dilution 1:400, #13120; Cell Signaling Technology), anti‐ MMP‐9 (rabbit polyclonal, dilution 1:1000, #ab38898; abcam), anti‐ MMP‐2 (rabbit polyclonal, dilution 1:1000, #ab97779; abcam), anti‐IL1RL2 (rabbit polyclonal, dilution 1:1000, # PA5‐38013; Invitrogen), anti‐p38 (rabbit polyclonal, dilution 1:5000, #9212; Cell signaling technology), anti‐phospho‐p38 (p‐p38) (rabbit polyclonal, dilution 1:1000, #9211; Cell signaling technology), anti‐JNK (rabbit polyclonal, dilution 1:5000, #9252; Cell signaling technology) and anti‐phospho‐JNK (p‐JNK) (rabbit polyclonal, dilution 1:1000, #9251; Cell signaling technology).

Techniques: Activation Assay, Western Blot, MANN-WHITNEY, Staining

Journal: bioRxiv

Article Title: EVIDENCE FOR ANGIOTENSIN II AS A NATURALLY EXISTING SUPPRESSOR FOR THE NATRIURETIC PEPTIDE SYSTEM

doi: 10.1101/2023.01.26.525806

Figure Lengend Snippet: (A) Protein expression of different human PKC isoforms in the membrane and cytosol fractions of different HEK293 transfected cell lines (with or without treatment of ANGII). GC-A and NaK-ATPase serve as quality control for the separation of membrane and cytosol fractions. GAPDH serves as loading control. (B) Protein expression of phosphorylated PKC substrates, phosphorylated p38 MAPK, and p38 MAPK in HEK293/GC-A + /AT 1 + cells treated with different PKC modulators. GAPDH serves as loading control. PMA, phorbol 12-myristate 13-acetate. (C) In vitro cGMP generation in HEK293/GC-A + /AT 1 + in response to ANP (10 −8 M) with or without ANGII (10 −8 M) and valsartan (1 μM) or Go6983 (5 μM). Values of cGMP were normalized to vehicle group (blue) under ANP treatment. * indicates P <0.05, one-way ANOVA with Dunnett multiple comparisons test. (D) Hypothetical mechanism underlying the crosstalk between ANGII and GC-A derived from our in vitro studies.

Article Snippet: The following primary antibodies were used: anti-GCA (catalog#MAB48601, R&D System, Minneapolis, MN) at 1:1000 dilution, anti-GCB (catalog#55113-1-AP, Proteintech, Rosemont, IL) at 1:200 dilution, anti-GFP (catalog#TA150041, OriGene, Rockville, MD) at 1:1000 dilution, anti-PKCα (catalog#2056, Cell Signaling Technology, Danvers, MA) at 1:500 dilution, anti-PKCε (catalog#2683, Cell Signaling Technology, Danvers, MA) at 1:1000 dilution, anti-Phspho-(Ser) PKC Substrate (catalog#2261, Cell Signaling Technology, Danvers, MA) at 1:1000 dilution, anti-NaK-ATPase (catalog#EP1845Y, abcam, Waltham, MA) at 1:2000 dilution, anti-p38 MAPK (catalog#8690, Cell Signaling Technology, Danvers, MA) at 1:1000 dilution, anti-Phospho-p38 MAPK (catalog#4511, Cell Signaling Technology, Danvers, MA) at 1:1000 dilution, and anti-GAPDH (catalog#2118, Cell Signaling Technology, Danvers, MA) at 1:5000 dilution.

Techniques: Expressing, Transfection, In Vitro, Derivative Assay

Journal: PLOS Neglected Tropical Diseases

Article Title: Clonorchis sinensis aggravates biliary fibrosis through promoting IL-6 production via toll-like receptor 2-mediated AKT and p38 signal pathways

doi: 10.1371/journal.pntd.0011062

Figure Lengend Snippet: Details of the antibodies used in this study.

Article Snippet: Phospho-p38 , Rabbit monoclonal , Rabbit IgG , 1:1000 , Cell Signaling Technology.

Techniques:

Journal: PLOS Neglected Tropical Diseases

Article Title: Clonorchis sinensis aggravates biliary fibrosis through promoting IL-6 production via toll-like receptor 2-mediated AKT and p38 signal pathways

doi: 10.1371/journal.pntd.0011062

Figure Lengend Snippet: (A, B) The phosphorylation of AKT, p65, ERK and p38 in liver were analyzed by Western blot. (C, D) 3 × 10 5 WT and TLR2 -/- mouse BECs were cultivated with C . sinensis ESPs (50 μg/mL) for 120 min, the cells were lysed in cells lysates and the phosphorylation of AKT, p65, ERK and p38 were analyzed by Western blot. (E) The production levels of IL-6 in the supernatant of BECs pretreated with or without AKT inhibitor, p38 inhibitor and ERK inhibitor for 1 h, then co-cultured with ESPs for 18 h were measured by ELISA.

Article Snippet: Phospho-p38 , Rabbit monoclonal , Rabbit IgG , 1:1000 , Cell Signaling Technology.

Techniques: Western Blot, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: PLOS Neglected Tropical Diseases

Article Title: Clonorchis sinensis aggravates biliary fibrosis through promoting IL-6 production via toll-like receptor 2-mediated AKT and p38 signal pathways

doi: 10.1371/journal.pntd.0011062

Figure Lengend Snippet: C . sinensis triggered AKT and p38 signal pathways dependent on TLR2 of BECs to promote the production of IL-6, which subsequently activated TGF-β1-Smad2/Smad3 pathways, myofibroblasts and up-regulated the expression of ECMs, finally resulting in biliary fibrosis in mice. In the figure, ‘ ↑ ’indicates positive regulation.

Article Snippet: Phospho-p38 , Rabbit monoclonal , Rabbit IgG , 1:1000 , Cell Signaling Technology.

Techniques: Expressing