Journal: Phytomedicine : international journal of phytotherapy and phytopharmacology

Article Title: Herbal formula Jiawei Xiaochengqi decoction prevents postoperative abdominal adhesion in a rat model through inhibition of CXCL2-CXCR2 pathway.

doi: 10.1016/j.phymed.2023.154662

Figure Lengend Snippet: Fig. 6. The effects of JWXCQ on expression of p-STAT3 and p-AKT in rats. Data were presented as the mean ± SD. *p<0.05, **p<0.01 and ***p<0.001 vs model group.

Article Snippet: Blots were reacted with the specific primary antibodies: phospho-STAT3 (Ser727) and phospho-AKT (Ser473) (WLP2412, WLP001a, Wanleibio, Shenyang, China), Collagen-1 (COL-1, 14695-1-AP, Proteintech, Chicago, USA), CXCR2 (A3301, ABclonal Technology, Wuhan, China), GAPDH (bs-0755R, Bioss Biotechnology, Beijing, China) and β-actin (81115-1-RR, Proteintech, Chicago, USA), followed by crosslinking with the secondary antibodies.

Techniques: Expressing

Journal: Phytomedicine : international journal of phytotherapy and phytopharmacology

Article Title: Herbal formula Jiawei Xiaochengqi decoction prevents postoperative abdominal adhesion in a rat model through inhibition of CXCL2-CXCR2 pathway.

doi: 10.1016/j.phymed.2023.154662

Figure Lengend Snippet: Fig. 9. The effects of JWXCQ on expression of CXCL2, CXCR2, p-STAT3 and p-AKT in cell experiments. (A) The levels of CXCL2 in Raw 264.7 macrophages su pernatant. (B-C) The protein expression of CXCR2 in Raw 264.7 macrophages and PF (primary fibroblasts from peritoneal adhesion tissues). (D-E) The protein expression of p-STAT3 and p-AKT in Raw 264.7 macrophages. (F-G) The protein expression of p-STAT3 and p-AKT in PF. Data were presented as the mean ± SD. *p<0.05, **p<0.01 and ***p<0.001 vs model or PF group.

Article Snippet: Blots were reacted with the specific primary antibodies: phospho-STAT3 (Ser727) and phospho-AKT (Ser473) (WLP2412, WLP001a, Wanleibio, Shenyang, China), Collagen-1 (COL-1, 14695-1-AP, Proteintech, Chicago, USA), CXCR2 (A3301, ABclonal Technology, Wuhan, China), GAPDH (bs-0755R, Bioss Biotechnology, Beijing, China) and β-actin (81115-1-RR, Proteintech, Chicago, USA), followed by crosslinking with the secondary antibodies.

Techniques: Expressing

Journal: Phytomedicine : international journal of phytotherapy and phytopharmacology

Article Title: Herbal formula Jiawei Xiaochengqi decoction prevents postoperative abdominal adhesion in a rat model through inhibition of CXCL2-CXCR2 pathway.

doi: 10.1016/j.phymed.2023.154662

Figure Lengend Snippet: Fig. 10. The effects of JWXCQ on expression of CXCR2, COL-1, p-STAT3 and p-AKT in siCXCR2 transfected PF (primary fibroblasts from peritoneal adhesion tissues). (A) The protein expression of CXCR2 in siCXCR2 transfected PF. (B-D) The protein expression of COL-1, p-STAT3 and p-AKT in siCXCR2 transfected PF. Data were presented as the mean ± SD. *p<0.05, **p<0.01 and ***p<0.001 vs PF group.

Article Snippet: Blots were reacted with the specific primary antibodies: phospho-STAT3 (Ser727) and phospho-AKT (Ser473) (WLP2412, WLP001a, Wanleibio, Shenyang, China), Collagen-1 (COL-1, 14695-1-AP, Proteintech, Chicago, USA), CXCR2 (A3301, ABclonal Technology, Wuhan, China), GAPDH (bs-0755R, Bioss Biotechnology, Beijing, China) and β-actin (81115-1-RR, Proteintech, Chicago, USA), followed by crosslinking with the secondary antibodies.

Techniques: Expressing, Transfection

Journal: Nucleic Acids Research

Article Title: Precise and efficient C-to-U RNA base editing with SNAP-CDAR-S

doi: 10.1093/nar/gkad598

Figure Lengend Snippet: Benchmark with Cas13-RESCUE-S on endogenous targets and applications. ( A ) Comparison of editing yields at various sites on various endogenous targets and one disease-relevant cDNA (APOE) comparing SNAP-CDAR-S with standard guide RNAs (30 nt, 6-C-23, 2′OMe gapmer) versus Cas13 RESCUE-S with plasmid-borne optimized Cas guide RNAs. Both editing enzymes were expressed from the same single genomic locus. ( B ) Comparing both tools, SNAP-CADR-S versus Cas RESCUE-S, for the activation of β-catenin by RNA editing. Given are C-to-U editing yields (T41I) and the luminescence-based read-out of pathway activation. For further controls, see . ( C ) Editing of the regulatory phospho-site serine 727-to-glycine in STAT3, read-out of editing yield by Sanger sequencing, and amount of total STAT3 and pS727 STAT3 protein by western blot. Data in (A), (B) and (C) are shown as the mean ± s.d. of N = 3 independent experiments.

Article Snippet: 30 μg of total protein was run on a NovexTM WedgeWellTM 8 to 16%, Tris-glycine, 1.0 mm, Mini Protein Gel (ThermoFisher Scientific) with 200 V for 60 min. Blotting was performed with a Mini Trans-Blot Cell ® (BioRad) at 100 V for 60 min. For protein detection, membranes were incubated with monoclonal anti-β-actin antibody produced in mouse (Sigma, 1:5000 dil.) and either Stat3 (DRZ2G) Rabbit mAb (CellSignaling, 1:1000 dil.) or P-Stat3 (S727) (D8C2Z) Rabbit mAb (CellSignaling, 1:1000 dil.).

Techniques: Comparison, Plasmid Preparation, Activation Assay, Sequencing, Western Blot

Journal: Oncotarget

Article Title: Impaired lymphatic function accelerates cancer growth

doi: 10.18632/oncotarget.9953

Figure Lengend Snippet: A. Multiplex ELISA of tumor interstitial fluid from C3HBA tumors demonstrating significantly higher IL-6 level in Chy compared to wt mice. Bars depict the mean cytokine concentration ± SEM, n=8 per group (except GM-CSF: n=3 per group). *p<0.05. B. ELISA of tumor interstitial fluid from KHT-1 tumors demonstrating no significant difference in IL-6 levels between Chy and wt mice. Bars depict the mean cytokine concentration ± SEM, n=5 per group. C. Tumor growth of C3HBA breast cancer in Chy and wt mice treated with either IL-6 receptor antibody (tocilizumab) or placebo (saline) i.p for 21 days. The graphs depict the mean tumor volume ± SEM, n=7 per group. Arrows indicate when the treatment started. Tocilizumab significantly reduced tumor growth both in Chy and wt mice. *p<0.05. D. Western blot analysis demonstrating reduced STAT3 phosphorylation (Ser727) in C3HBA tumors when IL-6 signaling is inhibited with tocilizumab. Densitometry of western blots presented as the ratio of protein of interest normalized to STAT3 and actin expression. Bars depict the mean ± SEM, n=3 per group. *p<0.05

Article Snippet: Rabbit anti-actin (Sigma), rat anti-IL1-beta/IL-1F2 (R&D), rabbit anti-TGF-beta (Nordic Biosite), rat anti-GM-CSF (Abcam), rat anti-IL10 (Abcam), goat anti-M-CSF (Abcam), goat anti-VEGFD (Abcam), rabbit anti-VEGF (Abcam), rabbit anti-VEGFC (Abcam), rabbit anti-PlGF (Abcam), rabbit anti-IL-6 (Abcam), rabbit anti-phospho (Ser727) STAT3 (Cell Signaling), rabbit anti-phospho, (Tyr705) STAT3 (Cell Signaling), rabbit anti-STAT3 (Cell Signaling).

Techniques: Multiplex Assay, Enzyme-linked Immunosorbent Assay, Concentration Assay, Saline, Western Blot, Phospho-proteomics, Expressing

Journal: BMJ Open Diabetes Research & Care

Article Title: Anti-inflammatory, antioxidant and renoprotective effects of SOCS1 mimetic peptide in the BTBR ob/ob mouse model of type 2 diabetes

doi: 10.1136/bmjdrc-2020-001242

Figure Lengend Snippet: JAK/STAT, inflammatory and oxidative stress pathways in kidney tissue of BTBR ob/ob model. (A) Representative images of phosphorylated (p-)STAT3, p-p65 NF-κB and p-NRF2, and quantification of positive cells in glomerular and tubular fields of BTBR WT and ob/ob mice. Magnification ×400 and ×630. Arrows indicate positive stained cells. (B) Representative images of SOCS1/SOCS3 proteins and quantification of positive stained area per tubular field. Magnification ×200. (C) Real-time PCR analysis of JAK/STAT, inflammatory and oxidative stress genes. Values normalized by endogenous control gene 18s are expressed as n-fold of the average value from BTBR WT. Data are shown as scatter dot plots and mean±SD of each group (n=5–7 mice/group); *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001 versus BTBR WT control. (D) Schematic protein–protein interaction prediction of JAK/STAT, inflammatory and oxidative stress markers in Mus musculus according to STRING software. More information can be found at https://string-db.org/ . a.u., arbitrary units; BTBR, black and tan brachyuric; GCS, glomerular cross section; JAK/STAT, Janus kinase/signal transducers and activators of transcription; NF-κB, nuclear factor-κB; NRF2, nuclear factor erythroid 2-related factor 2; ob/ob, obese/obese; SOCS, suppressor of cytokine signaling; WT, wild type.

Article Snippet: The primary antibodies for immunodetection were sourced as follows: phosphorylated (p-) STAT3 serine 727 (Cell Signaling Technology Cat# 9134, RRID:AB_331589, dilution 1:100), p-STAT1 tyrosine 701 (Cell Signaling Technology Cat# 7649, RRID:AB_10950970, dilution 1:50), p-p65 subunit of nuclear factor-κB (NF-κB) serine 536 (Santa Cruz Biotechnology Cat# sc-33020, RRID:AB_2179018, dilution 1:100), p-nuclear factor erythroid 2-related factor 2 (NRF2) serine 40 (Abcam Cat# ab76026, RRID:AB_1524049, dilution 1:2000), SOCS1 (Abcam Cat# ab62584, RRID:AB_956316, dilution 1:1000), SOCS3 (Abcam Cat# ab16030, RRID:AB_443287, dilution 1:200), F4/80 monocytes/macrophages (Bio-Rad Cat# MCA497, RRID:AB_2098196, dilution 1:70), CD3 T lymphocytes (Agilent Cat# M7254, RRID:AB_2631163, dilution 1:100), Wilms tumor protein-1 (WT-1; Agilent Cat# M3561, RRID:AB_2304486, dilution 1:100), perilipin-1 (sc-390169, dilution 1:50, Santa Cruz Biotechnology, USA) and 4-hydroxy-2-nonenal (4-HNE; Abcam Cat# ab46545, RRID:AB_722490, dilution 1:200).

Techniques: Staining, Real-time Polymerase Chain Reaction, Control, Software

Journal: BMJ Open Diabetes Research & Care

Article Title: Anti-inflammatory, antioxidant and renoprotective effects of SOCS1 mimetic peptide in the BTBR ob/ob mouse model of type 2 diabetes

doi: 10.1136/bmjdrc-2020-001242

Figure Lengend Snippet: MiS1 treatment inhibits kidney JAK/STAT activation and renal microinflammatory milieu in the BTBR ob/ob model. (A) Graphs and images represent the changes observed in JAK/STAT activation (p-STATs) in diabetic mice treated with active MiS1 (2 µg and 4 µg) and inactive mutated peptide (Mut 4 µg) compared with vehicle controls (Veh), quantified per number of positive cells p-STAT1+ and p-STAT3+, both at the glomerular and tubular fields. Magnification ×630. (B) Representative images of immunohistochemistry against F4/80 and CD3. Magnification ×200 and ×630. Graphs represent the quantification of average number of monocytes/macrophages F4/80+ and CD3+ T lymphocytes, both at the glomerular and interstitial fields. Arrows indicate positively stained cells. (C) Gene expression analysis of mRNA related with JAK/STAT pathway ( Stat1 , Stat3 , Socs1 and Socs3 ), inflammatory cytokines ( Tnfα and Il-12 ) and chemokines ( Mcp-1 and Rantes ), and kidney damage markers ( Kim-1 and Ngal ) were evaluated by real-time PCR, being normalized in each sample by endogenous control gene 18s and expressed as n-fold the average value obtained in the vehicle group (Veh). Data are shown as scatter dot plots and mean±SD of each group (n=6–7 mice/group); *p<0.05, **p<0.01, ****p<0.0001 versus diabetic vehicle control. BTBR, black and tan brachyuric; GCS, glomerular cross section; JAK/STAT, Janus kinase/signal transducers and activators of transcription; ob/ob, obese/obese.

Article Snippet: The primary antibodies for immunodetection were sourced as follows: phosphorylated (p-) STAT3 serine 727 (Cell Signaling Technology Cat# 9134, RRID:AB_331589, dilution 1:100), p-STAT1 tyrosine 701 (Cell Signaling Technology Cat# 7649, RRID:AB_10950970, dilution 1:50), p-p65 subunit of nuclear factor-κB (NF-κB) serine 536 (Santa Cruz Biotechnology Cat# sc-33020, RRID:AB_2179018, dilution 1:100), p-nuclear factor erythroid 2-related factor 2 (NRF2) serine 40 (Abcam Cat# ab76026, RRID:AB_1524049, dilution 1:2000), SOCS1 (Abcam Cat# ab62584, RRID:AB_956316, dilution 1:1000), SOCS3 (Abcam Cat# ab16030, RRID:AB_443287, dilution 1:200), F4/80 monocytes/macrophages (Bio-Rad Cat# MCA497, RRID:AB_2098196, dilution 1:70), CD3 T lymphocytes (Agilent Cat# M7254, RRID:AB_2631163, dilution 1:100), Wilms tumor protein-1 (WT-1; Agilent Cat# M3561, RRID:AB_2304486, dilution 1:100), perilipin-1 (sc-390169, dilution 1:50, Santa Cruz Biotechnology, USA) and 4-hydroxy-2-nonenal (4-HNE; Abcam Cat# ab46545, RRID:AB_722490, dilution 1:200).

Techniques: Activation Assay, Immunohistochemistry, Staining, Gene Expression, Real-time Polymerase Chain Reaction, Control

Journal: Oncotarget

Article Title: A novel small-molecule compound targeting CD147 inhibits the motility and invasion of hepatocellular carcinoma cells

doi: 10.18632/oncotarget.6990

Figure Lengend Snippet: A. The effect of AC-73 on MMP-2 mRNA level in SMMC-7721 cells. MMP-2 mRNA levels were detected using real-time PCR after treatment with DMSO or different concentrations of AC-73. Here, 20 μg/ml HAb18 was used as a positive control. B. Gelatin zymography analysis of MMP-2 secretion. Top, representative image; bottom, quantification of the grayscale analysis in three independent experiments. C, D. CD147, ERK1/2 and STAT3 activities and total ERK1/2 and STAT3 expression were examined in SMMC-7721 cells treated with different concentrations of AC-73. E, F. ERK1/2 and STAT3 activities and total ERK1/2 and STAT3 expression were examined in SMMC-7721 cells with different treatments. AC-73 was used at a concentration of 10 μM, the ERK1/2 inhibitor PD0325901 was used at 1 μM, and the STAT3 inhibitor WP1066 was used at 10 μM. G, H. ERK1/2 and STAT3 activities were examined in total lysates of HCC and HCC CD147 −/− cells using Western blotting. All the Histograms show quantification of the gray scale analysis for western blotting. The bars represent each sample performed in triplicate and the error bars indicate ± SD. (*** P < 0.001, ** P < 0.01, * P < 0.05, # P > 0.05), one-way ANOVA (H) for multiple comparisons (A, B, D and F), Student's t -test for two comparisons (H).

Article Snippet: Antibodies against STAT3 and p-STAT3 (Ser727) were obtained from Biorbyt LLC (San Francisco, CA).

Techniques: Real-time Polymerase Chain Reaction, Positive Control, Zymography, Expressing, Concentration Assay, Western Blot

Journal: Oncotarget

Article Title: A novel small-molecule compound targeting CD147 inhibits the motility and invasion of hepatocellular carcinoma cells

doi: 10.18632/oncotarget.6990

Figure Lengend Snippet: A, B. Metastasis assay by intrahepatic injection of SMMC-7721 cells in nude mice. Liver were excised for examination, scale bars: 1cm. Intrahepatic metastases in each group, as determined by calculation of metastatic lesions in the whole liver ( n = 5). C, D. Phosphorylation of ERK1/2 and STAT3 was examined in total lysates of original orthotopic tumors from each group, and histogram shows quantification of the gray scale analysis for western blotting. E. Phosphorylation of ERK1/2 and STAT3, MMP-2 and CD147 expression in original orthotopic tumors at 200× magnification, as determined by IHC staining analysis following treatment with AC-73. Each inset shows images obtained at 400× magnification, scale bars: 100 μm.

Article Snippet: Antibodies against STAT3 and p-STAT3 (Ser727) were obtained from Biorbyt LLC (San Francisco, CA).

Techniques: Injection, Phospho-proteomics, Western Blot, Expressing, Immunohistochemistry

Journal: Oncotarget

Article Title: A novel small-molecule compound targeting CD147 inhibits the motility and invasion of hepatocellular carcinoma cells

doi: 10.18632/oncotarget.6990

Figure Lengend Snippet: A proposed working model for AC-73-mediated suppression of CD147/ERK1/2/STAT3 signaling to inhibit HCC metastasis

Article Snippet: Antibodies against STAT3 and p-STAT3 (Ser727) were obtained from Biorbyt LLC (San Francisco, CA).

Techniques:

Journal: Disease Markers

Article Title: Anti-inflammatory Effects of Alcohol Are Associated with JNK-STAT3 Downregulation in an In Vitro Inflammation Model in HepG2 Cells

doi: 10.1155/2021/6622701

Figure Lengend Snippet: Protein expression levels of phosphorylated SAPK/JNK (Thr183/Tyr185, (a) or phosphorylated STAT3 (Ser727, (b) levels in HepG2 cells are shown. Cells were stimulated with IL-1 β (1 ng/ml) and treated with low (LoD, 85 mM)- or high (HiD, 170 mM)-dose EtOH for 1 h (acute) or 72 h (prolonged). Unstimulated controls (-); IL-1 β -stimulated samples (+). p < 0.05: ∗ vs . unstimulated controls, # vs . stimulated and EtOH-untreated controls.

Article Snippet: The FastScanTM Phospho-Stat3 (Ser727) ELISA Kit is a sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of Stat3 when phosphorylated at Ser727 (Cell Signaling Technology).

Techniques: Expressing