Journal: Journal of Translational Medicine

Article Title: IL-27 attenuates airway inflammation in a mouse asthma model via the STAT1 and GADD45γ/p38 MAPK pathways

doi: 10.1186/s12967-016-1039-x

Figure Lengend Snippet: The STAT1 and GADD45γ/p38 MAPK pathways remain impairment in the IL-27 treatment groups. a Protocol of OVA-induced allergic asthma and IL-27 administration. b The mRNA expression of STAT1, GADD45-γ and p38 MAPK by qPCR. c The protein expression of STAT1, GADD45γ and p38 MAPK by Western blot. In IL-27 treated group, p-STAT1, p-p38MAPK were still impaired compared with PBS group, while GADD45γ had no markedly difference. d Densitometer measurements of pSTAT1/GAPDH, GADD45γ/GAPDH and p38 MAPK/GAPDH. IL-27 treatment had no improvement in the already impaired signal pathway.(GAPDH expression was acted as internal reference.) The columns and error bars represent the mean and SEM (n = 6 per group). *P < 0.05, **P < 0.01, ***P < 0.001. Similar results were obtained in at least three independent experiments

Article Snippet: The following antibodies were also used: anti-STAT1 antibody (SC592; Santa Cruz Biotechnology, Santa Cruz, CA, USA); anti-Py-STAT1 antibody (catalog no. 9171; Cell Signaling, Boston, MA, USA), anti-GADD45γ antibody (ab196774,Abcam,Cambridge,UK), anti-p38 MAPK antibody (CST9212, Cell Signaling, Boston, MA, USA), and anti-phospho-p38 MAPK antibody (CST9215, Cell Signaling, Boston, MA, USA).

Techniques: Expressing, Western Blot

Journal: Journal of Translational Medicine

Article Title: IL-27 attenuates airway inflammation in a mouse asthma model via the STAT1 and GADD45γ/p38 MAPK pathways

doi: 10.1186/s12967-016-1039-x

Figure Lengend Snippet: The STAT1 and GADD45γ/p38 MAPK pathways are up-regulated in the IL-27 prevention groups. a Protocol of OVA-induced allergic asthma and IL-27 administration. b The mRNA expression of STAT1, GADD45-γ and p38 MAPK by qPCR. It showed IL-27 preventative treatment reverse the impaired STAT1 and strengthen GADD45-γ expression as well. c The protein expression of STAT1, GADD45-γ and p38 MAPK by Western blot. It shows IL-27 preventative treatment reverse the impaired p-STAT1 and strengthen GADD45-γ/p38 MAPK protein expression as well. d Densitometer measurements of pSTAT1/GAPDH, GADD45-γ/GAPDH and p38 MAPK/GAPDH. GAPDH expression was acted as internal reference. The columns and error bars represent the mean and SEM (n = 6 per group). *P < 0.05, **P < 0.01, ***P < 0.001. Similar results were obtained in at least three independent experiments

Article Snippet: The following antibodies were also used: anti-STAT1 antibody (SC592; Santa Cruz Biotechnology, Santa Cruz, CA, USA); anti-Py-STAT1 antibody (catalog no. 9171; Cell Signaling, Boston, MA, USA), anti-GADD45γ antibody (ab196774,Abcam,Cambridge,UK), anti-p38 MAPK antibody (CST9212, Cell Signaling, Boston, MA, USA), and anti-phospho-p38 MAPK antibody (CST9215, Cell Signaling, Boston, MA, USA).

Techniques: Expressing, Western Blot

Journal: Clinical and Molecular Hepatology

Article Title: Interferon-free treatment for hepatitis C virus infection induces normalization of extrahepatic type I interferon signaling

doi: 10.3350/cmh.2017.0074

Figure Lengend Snippet: STAT1 phosphorylation decreased in the peripheral blood mononuclear cells (PBMCs) of HCV-infected patients after direct-acting antiviral (DAA) treatment. (A, C) PBMCs from HCV-infected patients were isolated before DAA (pre DAA) and at the end of DAA (post DAA) treatment. Immunoblotting was performed to detect the protein levels of STAT1, PY-STAT1, and GAPDH. (B) PBMCs from HCV-infected patients were isolated before DAA treatment. TaqMan real-time quantitative polymerase chain reaction was performed to detect mRNA levels of IFI44, CXCL10, and GAPDH. Pearson's correlation analysis was performed to identify the associations between expression levels of IFI44/CXCL10 and relative band intensity of PY-STAT1. STAT1, signal transducer and activator of transcription 1; HCV, hepatitis C virus; PY-STAT1, tyrosine-phosphorylated STAT1; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; IFI44, interferon-induced protein 44; CXCL10, C-X-C motif chemokine ligand 10. * P <0.05.

Article Snippet: The antibodies used are as follows: STAT1 (1:1,000, rabbit, BD transduction laboratories, San Jose, CA, USA), PY-STAT1 (1:1,000, mouse, BD transduction laboratories), GAPDH (1:1,000, rabbit, Santa Cruz Biotechnology, Santa Cruz, CA, USA), HRP-conjugated rabbit immunoglobulin G (IgG) (1:5000, Santa Cruz Biotechnology, Santa Cruz, CA, USA), and HRP-conjugated mouse IgG (1:5,000, Santa Cruz Biotechnology, Santa Cruz, CA, USA).

Techniques: Infection, Isolation, Western Blot, Real-time Polymerase Chain Reaction, Expressing

Journal: Journal of Advanced Research

Article Title: A novel TGR5 agonist Sauchinone ameliorates IMQ induced murine psoriasis by regulating macrophage polarization

doi: 10.1016/j.jare.2025.04.034

Figure Lengend Snippet: Sauchinone inhibits M1 macrophage polarization and promotes M2 macrophage polarization in BMDM. (A) The cytotoxicity of Sauchinone on BMDM was detected by CCK-8 kit. (B-G) BMDM were stimulated with LPS plus IFN-γ to induce M1 polarization in the presence or absence of different doses of Sauchinone. The transcription levels of Tnf-α , Il-6 , Il −12 , Inos , Il-1β , Cox-2 and Nlrp3 (B) were detected by RT-qPCR. TNF-α, IL-6, IL-12 and NO levels in culture supernatants (C) were measured by ELISA or Griess assay. The protein levels of p-STAT-1, STAT-1, and INOS in cell lysates (D) were detected by western blot. The percentages of INOS + , MHCⅡ + and CD86 + BMDM (E) were measured by FACS. (F-H) BMDM were stimulated with IL-4 plus IL-13 to induce M2 polarization in the presence or absence of different doses of Sauchinone. The transcription levels of Arg-1, Fizz1 , Ym-1 , Cd206 , Cd301 , and Dectin-1 (F) were detected by RT-qPCR. The protein levels of p-STAT-6 and STAT-6 in cell lysates (G) were detected by western blot. The percentages of CD301 + BMDM (H) were measured by FACS. Data were shown as Mean ± SEM from triplicate measurements. *P < 0.05, **P < 0.01 compared as indicated.

Article Snippet: Anti-mouse STAT-1 (9712L), anti-mouse p-STAT-1 (7649L), anti-mouse Inducible nitric oxide synthase (INOS) (13120S), Anti-mouse STAT-6 (9362S), anti-mouse p-STAT-6 (56554S), anti-mouse PKA (5842S), anti-mouse p-CREB (9197S), anti-mouse CREB (9198L),anti-mouse p-IκB (9246L), anti-mouse IκB (4812S), anti-mouse p-p65 (3033L), anti-mouse p65 (4764S), anti-mouse ASC (67824S) and anti-mouse NLRP3 (15101S) were obtained from Cell Signaling Technology (Danvers, MA, USA).

Techniques: CCK-8 Assay, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Griess Assay, Western Blot

Journal: Journal of Advanced Research

Article Title: A novel TGR5 agonist Sauchinone ameliorates IMQ induced murine psoriasis by regulating macrophage polarization

doi: 10.1016/j.jare.2025.04.034

Figure Lengend Snippet: Sauchinone inhibits macrophage M1 polarization in TGR5 dependent manner. (A-B) BMDM from WT mice were stimulated with LPS plus IFN-γ to induce M1 polarization in the presence or absence of Sauchinone (30 μM) or SBI-115 (5 μM). The transcription levels of Tnf-α , Il-6 , Inos and Il-1β (A) were detected by RT-qPCR. The protein levels of p-STAT-1, STAT-1 and INOS in cell lysates (B) were detected by western blot. (C-E) BMDM from WT mice were stimulated with LPS plus IFN-γ to induce M1 polarization in the presence or absence of Sauchinone (30 μM) or TGR5 si-RNA. The transcription levels of Tnf-α , Il-6 , Inos and Il-1β (C) were detected by RT-qPCR. The protein levels of p-STAT-1, STAT-1 and INOS in cell lysates (D) were detected by western blot. IL-6 and NO levels in culture supernatants (E) were measured by ELISA and Griess assay. (F-H) BMDM from Tgr5 -/- mice were stimulated with LPS plus IFN-γ to induce M1 polarization in the presence or absence of Sauchinone (30 μM). IL-6 and NO levels in culture supernatants (F) were measured by ELISA or Griess assay. The transcription levels of Tnf-α , Il-6 , Inos and Il-1β (G) were detected by RT-qPCR. The protein levels of p-STAT-1, STAT-1 and INOS in cell lysates (H) were detected by western blot. Data were shown as Mean ± SEM from triplicate measurements. *P < 0.05, **P < 0.01 compared as indicated.

Article Snippet: Anti-mouse STAT-1 (9712L), anti-mouse p-STAT-1 (7649L), anti-mouse Inducible nitric oxide synthase (INOS) (13120S), Anti-mouse STAT-6 (9362S), anti-mouse p-STAT-6 (56554S), anti-mouse PKA (5842S), anti-mouse p-CREB (9197S), anti-mouse CREB (9198L),anti-mouse p-IκB (9246L), anti-mouse IκB (4812S), anti-mouse p-p65 (3033L), anti-mouse p65 (4764S), anti-mouse ASC (67824S) and anti-mouse NLRP3 (15101S) were obtained from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Quantitative RT-PCR, Western Blot, Enzyme-linked Immunosorbent Assay, Griess Assay

Journal: Journal of Advanced Research

Article Title: A novel TGR5 agonist Sauchinone ameliorates IMQ induced murine psoriasis by regulating macrophage polarization

doi: 10.1016/j.jare.2025.04.034

Figure Lengend Snippet: Sauchinone inhibits macrophage M1 polarization through TGR5/PKA pathway. (A-C) BMDM were stimulated with LPS plus IFN-γ to induce M1 polarization in the presence or absence of different doses of Sauchinone (30 μM) or PKA inhibitor H89 (10 μM). IL-6 and NO levels in culture supernatants (A) were measured by ELISA or Griess assay. The transcription levels of Tnf-α , Il-6 , Inos and Il-1β (B) were detected by RT-qPCR. The protein levels of p-STAT-1, STAT-1 and INOS in cell lysates (C) were detected by western blot. Data were shown as Mean ± SEM from triplicate measurements. *P < 0.05, **P < 0.01 compared as indicated.

Article Snippet: Anti-mouse STAT-1 (9712L), anti-mouse p-STAT-1 (7649L), anti-mouse Inducible nitric oxide synthase (INOS) (13120S), Anti-mouse STAT-6 (9362S), anti-mouse p-STAT-6 (56554S), anti-mouse PKA (5842S), anti-mouse p-CREB (9197S), anti-mouse CREB (9198L),anti-mouse p-IκB (9246L), anti-mouse IκB (4812S), anti-mouse p-p65 (3033L), anti-mouse p65 (4764S), anti-mouse ASC (67824S) and anti-mouse NLRP3 (15101S) were obtained from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Enzyme-linked Immunosorbent Assay, Griess Assay, Quantitative RT-PCR, Western Blot

Journal: Journal of Advanced Research

Article Title: A novel TGR5 agonist Sauchinone ameliorates IMQ induced murine psoriasis by regulating macrophage polarization

doi: 10.1016/j.jare.2025.04.034

Figure Lengend Snippet: Sauchinone normalizes TGR5/PKA/CREB pathway and regulates M1/M2 macrophages markers in skin of IMQ-induced psoriasis mice. (A) The protein levels of TGR5, PKA. p-CREB, CREB in skin tissues were detected by western blot. (B) Cytokines including IL-6, IL-12 TNF-α, IFN-γ, IL-17 and IL-10 levels in serum were measured by ELISA (n = 6). (C) Cytokines including IL-6, IL-12, TNF-α, IFN-γ, IL-1β and IL-10 levels in skin homogenates were measured by ELISA (n = 6). (D) The mRNA levels of M2 markers including Ym-1 , Cd-301 , Dectin-1 , Arg-1 , Irf-4 and Ppar-γ in skin tissues were measured by qPCR (n = 6). (E) The protein levels of M1 markers including p-STAT-1, STAT1, p-p65, p65, NLRP3, p-IκB, IκB, Pro Caspase-1 and ASC in skin tissues were detected by western blot. Data were shown as Mean ± SEM from triplicate measurements. *P < 0.05, **P < 0.01 compared as indicated.

Article Snippet: Anti-mouse STAT-1 (9712L), anti-mouse p-STAT-1 (7649L), anti-mouse Inducible nitric oxide synthase (INOS) (13120S), Anti-mouse STAT-6 (9362S), anti-mouse p-STAT-6 (56554S), anti-mouse PKA (5842S), anti-mouse p-CREB (9197S), anti-mouse CREB (9198L),anti-mouse p-IκB (9246L), anti-mouse IκB (4812S), anti-mouse p-p65 (3033L), anti-mouse p65 (4764S), anti-mouse ASC (67824S) and anti-mouse NLRP3 (15101S) were obtained from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Western Blot, Enzyme-linked Immunosorbent Assay

Journal: CNS Neuroscience & Therapeutics

Article Title: CD8 + T cell infiltration and proliferation in the brainstem during experimental cerebral malaria

doi: 10.1111/cns.14431

Figure Lengend Snippet: Brainstem parenchyma cells responded to interferon signaling in experimental cerebral malaria (ECM) mice. (A) Violin plots showing the expression of IFN‐γ receptor 1 ( Ifngr1 ) in various brain parenchyma cell types both in the ECM and healthy mice. Each dot represents the gene expression levels in a single cell. (B) Expression of the Bst2 , Ifit1 , and Stat1 in the ECM brainstem and healthy brainstem. Each dot represents the gene expression levels in a single cell. *** p < 0.001. (C) Immunofluorescence staining of STAT‐1 and IBA‐1 in ECM brainstem and healthy brainstem. Red, STAT‐1; green, IBA‐1; blue, nuclei. The red arrow indicates STAT1‐positive microglia in the ECM brainstem. Scale bar, 10 μm. (D) Western blot results of p‐STAT‐1 from the ECM brainstem; samples were taken from ECM mice at 1, 3, 5, and 7 dpi. Data are mean ± SEM. Each dot represents one replicate, n = 3, ** p < 0.01. (E) Expression of the chemokines Cxcl9 , Cxcl10 , Ccl2 , and Ccl4 in microglia and astrocytes from ECM mice and healthy mice. Each dot represents the gene expression levels in a single cell. *** p < 0.001. (F) Expression of the chemokine receptors Cxcr3 , Ccr2 , and Ccr5 in various cell types from ECM brainstem parenchyma cells. Each dot represents the gene expression levels in a single cell. (G) Flow cytometry showing the expression of CXCR3 in brain‐infiltrated T cells and peripheral CD8 + T cells in ECM mice, three independent experiments.

Article Snippet: Antibodies against mouse PD‐L1 (Cat.66248), CD8a (Cat.65069), NeuN (Cat.26975), CCL‐2 (Cat.25542), CD18 (Cat.10554), and STAT‐1 (Cat.10144) were purchased from Proteintech.

Techniques: Expressing, Gene Expression, Immunofluorescence, Staining, Western Blot, Flow Cytometry