Journal: International Journal of Immunopathology and Pharmacology

Article Title: Protective effect of baicalin against arsenic trioxide-induced acute hepatic injury in mice through JAK2/STAT3 signaling pathway

doi: 10.1177/20587384211073397

Figure Lengend Snippet: Effects of BA on JAK2/STAT3 signaling pathway in ATO-induced mice. (a) Western blot analysis was used to assess the hepatic protein expression levels of JAK2, p-JAK2, STAT3, and p-STAT3. (b) The relative expression level of p-JAK2. (c) The relative expression level of p-STAT3. The results are rendered by the mean ± SEM ( n = 3). ## p < 0.01 compared with the CON group, * p < 0.05 and ** p < 0.01 compared with the ATO group. Abbreviations: CON: control group; ATO: arsenic trioxide-treated group; L-BA: low-dose baicalin group; H-BA: high-dose baicalin group; BA: baicalin alone group; SEM: standard error of the mean.

Article Snippet: Antibodies: anti-Bcl-2 associated X protein (BAX) (Servicebio, Wuhan; GB11690), anti-B-cell lymphoma 2 (Bcl-2) (Cloud-clonal, Wuhan; PAA778Mu01), anti-caspase-3 (Proteintech Group, Wuhan; 66,470-2-lg), anti-NF-κB (Servicebio, Wuhan; GB11142), anti-JAK2 (Servicebio, Wuhan; GB11325), anti-p-JAK2 (BIOSS, Beijing; BS-2485R), anti-STAT3 (Servicebio, Wuhan; GB11176), anti-p- STAT3 (Ruiying Biological, Jiangsu; RLP0250), and anti-β-actin (Servicebio, Wuhan; GB12001).

Techniques: Western Blot, Expressing

Journal: medRxiv

Article Title: Polymorphism in IFNAR contributes to glucocorticoid response and outcome in ARDS and COVID-19

doi: 10.1101/2022.03.10.22272123

Figure Lengend Snippet: (A ) STAT1 expression in the lung after 4-day culture in the presence of IFN beta with or without hydrocortisone (HC). ( B) pSTAT1 expression in the same specimens as in A. ( C) Example photomicrographs showing higher STAT2 expression in a TT patient than in a CT patient and the effect of HC on its nuclear translocation. Most STAT2 remains in the cytoplasm of the CT patients, whereas nuclear expression is prominent in the TT patient. Indicated insets are shown in the bottom row. Arrows. ( D ) Combined results of all patients noting that two CT samples are excluded in the data as the patients were already under glucocorticoid treatment at the time of sample acquisition. Ns, not significant; *P<0.05; **P<0.01; and ***P<0.001

Article Snippet: The first stage antibodies were anti-alpha chain of the IFN alpha/beta receptor (St John’s Laboratory STJ112765) that was used 1:2000 and 1:5000 and anti-Stat1 (1:400, 9175S), anti-pStat1(1:100, 9167S) and anti-Stat2 (1:200, 72604S) all from Cell Signalling.

Techniques: Expressing, Translocation Assay

Journal: Journal of Hepatocellular Carcinoma

Article Title: Blocking the CCL5–CCR5 Axis Using Maraviroc Promotes M1 Polarization of Macrophages Cocultured with Irradiated Hepatoma Cells

doi: 10.2147/JHC.S300165

Figure Lengend Snippet: The CCL5–CCR5 axis modulates macrophage polarization through the STAT3–SOCS3 pathway. ( A ) Representative Western blots showing the expression of the SOCS3 protein as well as total protein levels and phosphorylated-protein levels of JAK2, STAT3, p38 MAPK, ERK1/2, and cEBP/β in THP-1 M0 macrophages treated with CCL5 (0, 5, 10, or 20 ng/mL) for 24 h. ( B – D ) The most efficient siRNA against SOCS3 (labeled as si-SOCS3) #3 was determined by qRT-PCR ( B ) and Western blotting ( C ) and then transfected into THP-1 M0 macrophages. The markers of M1 (IL-12) and M2 (IL-10) macrophages were quantitated by qRT-PCR in transfected THP-1 M0 macrophages ( D ). ( E ) Representative Western blots revealing the expression of the SOCS3 protein as well as total protein levels and phosphorylated-protein levels of JAK2 and STAT3 in THP-1 M0 macrophages treated with CCL5 (5 ng/mL) and/or MVC (5 μM) for 24 h. ( F ) qRT-PCR analysis of the expression of TNF-α, IL-12, TGF-β1, and IL-10 in THP-1 M0 macrophages incubated with MVC and/or a STAT3 inhibitor (HJC0152). ( G ) Representative Western blots illustrating the expression of the SOCS3 protein as well as total protein levels and phosphorylated-protein levels of JAK2, STAT3, p38 MAPK, ERK1/2, and cEBP/β in THP-1 M0 macrophages cocultured with either HCCLM3 or Huh7 hepatoma cells subjected or not subjected to 8 Gy X-ray irradiation. The data are presented as mean ± standard deviation from three independent experiments. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: Antibodies against SOCS3 (cat. # ab16030), Janus kinase 2 (JAK2, ab108596), phosphorylated JAK2 (ab32101), STAT3 (ab68153), CCAAT/enhancer-binding protein (cEBP/β, ab32358), phosphorylated cEBP/β (ab52194), phosphorylated extracellular regulated protein kinases 1/2 (ERK1/2, ab76299), p38 mitogen-activated protein kinase (MAPK, ab170099), phosphorylated p38 MAPK (ab195049), IL-1β (ab156791), arginase 1 (Arg-1, ab239731), and CCL5 (ab9679) were bought from Abcam.

Techniques: Western Blot, Expressing, Labeling, Quantitative RT-PCR, Transfection, Incubation, Irradiation, Standard Deviation

Journal: Oncology reports

Article Title: Silibinin downregulates MMP2 expression via Jak2/STAT3 pathway and inhibits the migration and invasive potential in MDA-MB-231 cells.

doi: 10.3892/or.2017.5588

Figure Lengend Snippet: Figure 2. Silibinin downregulates the expression of Jak2/STAT3 signaling proteins in a dose- and time-dependent manner. (A) Western blot analyses showing the concentration dependent effect of silibinin in MDA‑MB‑231 cells following exposure to silibinin for 24 h. (B) Relative levels of the pSTAT3, STAT3, pJak2, and Jak2 proteins. (C) Time-dependent effect of silibinin on protein expression in MDA‑MB‑231 cells. (D) Relative expression levels of pSTAT3, STAT3, pJak2, and Jak2, measured using densitometry. These data were normalized to actin levels, and then shown as a percentage of the control. The data presented are representative of three independent experiments. Statistical analyses were conducted using the t-test (**p<0.01, ***p<0.001).

Article Snippet: An anti phosphor Jak2 antibody (Tyrosine residue 1007/1008) was purchased from Cell Signaling Technology (Beverly, MA, USA).

Techniques: Expressing, Western Blot, Concentration Assay, Control

Journal: Oncology reports

Article Title: Silibinin downregulates MMP2 expression via Jak2/STAT3 pathway and inhibits the migration and invasive potential in MDA-MB-231 cells.

doi: 10.3892/or.2017.5588

Figure Lengend Snippet: Figure 7. Schematic representation of the inhibition of invasive mechanisms provoked by silibinin in MDA‑MB‑231 cells. Silibinin inhibits Jak2 expression and phosphorylation, resulting, in turn, in the inhibition of STAT3 expression, phosphorylation, nuclear translocation, and DNA binding activity. Consequently, STAT3's down-stream targets are inhibited (including MMP2), resulting in reduced cell migration and invasion.

Article Snippet: An anti phosphor Jak2 antibody (Tyrosine residue 1007/1008) was purchased from Cell Signaling Technology (Beverly, MA, USA).

Techniques: Inhibition, Expressing, Phospho-proteomics, Translocation Assay, Binding Assay, Activity Assay, Migration

Journal: Molecular Medicine Reports

Article Title: Synergistic anticancer effects of ruxolitinib and calcitriol in estrogen receptor-positive, human epidermal growth factor receptor 2-positive breast cancer cells

doi: 10.3892/mmr.2018.8580

Figure Lengend Snippet: The expression of VDR and JAK2 in MCF7-HER18 breast cancer cells was demonstrated by western blot analysis. VDR, vitamin D receptor; JAK, Janus kinase; HER, human epidermal growth factor receptor.

Article Snippet: Antibodies against caspase-3 (cat. no. 9662S), apoptosis regulator Bcl-2 (Bcl-2; cat. no. 2876S), Bcl-2-like protein 1 (Bcl-xL; cat. no. 2762S), Bcl-2-associated agonist of cell death (BAD; cat. no. 9292S), cyclin-D1 (cat. no. 2922S), JAK2 (cat. no. 3230S), phosphorylated JAK2 (p-JAK2; cat. no. 3776S), c-Myc proto-oncogene protein (c-Myc; cat. no. 13987S) and β-actin (cat. no. 4967S) were purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA).

Techniques: Expressing, Western Blot

Journal: Molecular Medicine Reports

Article Title: Synergistic anticancer effects of ruxolitinib and calcitriol in estrogen receptor-positive, human epidermal growth factor receptor 2-positive breast cancer cells

doi: 10.3892/mmr.2018.8580

Figure Lengend Snippet: Levels of JAK2, p-JAK2, c-Myc, cyclin-D1, caspase 3, Bcl-2, Bcl-xL, BAD and β-actin proteins in MCF7-HER18 cells were determined by western blot analysis following the indicated 72 h treatments. The data are presented as the mean ± standard deviation. *P<0.05. JAK, Janus kinase; Bcl-2, apoptosis regulator Bcl-2; Bcl-xL, Bcl-2-like protein 1; BAD, Bcl-2-associated agonist of cell death; Myc, Myc proto oncogene protein HER, human epidermal growth factor receptor.

Article Snippet: Antibodies against caspase-3 (cat. no. 9662S), apoptosis regulator Bcl-2 (Bcl-2; cat. no. 2876S), Bcl-2-like protein 1 (Bcl-xL; cat. no. 2762S), Bcl-2-associated agonist of cell death (BAD; cat. no. 9292S), cyclin-D1 (cat. no. 2922S), JAK2 (cat. no. 3230S), phosphorylated JAK2 (p-JAK2; cat. no. 3776S), c-Myc proto-oncogene protein (c-Myc; cat. no. 13987S) and β-actin (cat. no. 4967S) were purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA).

Techniques: Western Blot, Standard Deviation

Journal: Journal of cardiovascular pharmacology

Article Title: Recombinant human erythropoietin suppresses endothelial cell apoptosis and reduces the ratio of Bax to Bcl-2 proteins in the aortas of apolipoprotein E-deficient mice

doi: 10.1097/FJC.0b013e31820d92fd

Figure Lengend Snippet: (A) Expression of EPOR in HUVECs treated with medium alone or EPO (5U/ml) for 24 hours as determined by Western blotting. K-562 lysate was used as a positive control. Data shown is representative of three independent experiments. (B) Expression of EPOR protein in murine aorta. Aortas were harvested from 10 individual mice and whole aortic lysate from each animal was subjected to Western blotting. These data depict 5 of the total of 10 aortic lysates analyzed in this experiment. (C) Expression of EPOR in murine aorta measured by qRT-PCR. These data represent mean ± s.e. of a total of 10 mice. (D) Expression of total Jak2, Stat5 and their phosphorylated forms, respectively, in HUVECs treated with either Medium or EPO for 24 hours as determined by Western blotting. Total HeLa cell extracts (prepared with our without interferon-α treatment and purchased from the manufacturer of the Jak 2 and Stat5 antibodies) were used as positive or negative controls.

Article Snippet: Antibodies against Jak2, Stat 5 and their phosphorylated forms (catalog numbers 3230S, 3776S, 9351 and 9363) were purchased from Cell Signaling Technology (Danvers, MA) and were used according to directions from the manufacturer.

Techniques: Expressing, Western Blot, Positive Control, Quantitative RT-PCR