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: Experimental Animals

Article Title: Danshen attenuates cartilage injuries in osteoarthritis in vivo and in vitro by activating JAK2/STAT3 and AKT pathways

doi: 10.1538/expanim.17-0062

Figure Lengend Snippet: Danshen attenuated articular cartilage degradation in vivo and activated the JAK2/STAT3 pathway. (A) Histological examination (H&E staining) of articular cartilage in Danshen- or SH-treated OA models. Scale bar=100 µ m. (B) Western blot was performed to assess the levels of phosphorylated and total JAK2 and STAT3 in articular cartilage.

Article Snippet: Primary antibodies against JAK2 (1:500; rabbit polyconal antibody, bs-23004R, Bioss, Beijing, China), phosphorylated JAK2 (p-JAK2) (1:500; rabbit polyclonal antibody, bs-2485R, Bioss), STAT3 (1:400; rabbit polyclonal antibody, bs-20382R, Bioss), p-STAT3 (1:500; rabbit polyclonal antibody, bs-3429R, Bioss), B-cell lymphoma 2 (Bcl-2) (1:500; rabbit polyclonal antibody, bs-0032R, Bioss), Bcl-2-associated X protein (Bax) (1:500; rabbit polyclonal antibody, bs-4564R, Bioss), cleaved caspase-3 (1:500; rabbit polyclonal antibody, WL01992, Wanleibio, Shenyang, China) and cleaved poly (ADP-ribose) polymerase (PARP) (1:500; rabbit polyclonal antibody, WL01932, Wanleibio), AKT (1:500; rabbit polyclonal antibody, bs-6951R, Bioss), or p-AKT (1:1,000; mouse monoclonal antibody, bsm-33281M, Bioss) were diluted in 5% skim milk.

Techniques: In Vivo, Staining, Western Blot

Journal: Experimental Animals

Article Title: Danshen attenuates cartilage injuries in osteoarthritis in vivo and in vitro by activating JAK2/STAT3 and AKT pathways

doi: 10.1538/expanim.17-0062

Figure Lengend Snippet: Danshen activated the JAK2/ STAT3 pathway. JAK and p-JAK2 (A) and STAT3 and p-STAT3 (B) were detected by western blot. The levels of p-JAK and p-STAT3 were upregulated by Danshen in SNP-treated chondrocytes.

Article Snippet: Primary antibodies against JAK2 (1:500; rabbit polyconal antibody, bs-23004R, Bioss, Beijing, China), phosphorylated JAK2 (p-JAK2) (1:500; rabbit polyclonal antibody, bs-2485R, Bioss), STAT3 (1:400; rabbit polyclonal antibody, bs-20382R, Bioss), p-STAT3 (1:500; rabbit polyclonal antibody, bs-3429R, Bioss), B-cell lymphoma 2 (Bcl-2) (1:500; rabbit polyclonal antibody, bs-0032R, Bioss), Bcl-2-associated X protein (Bax) (1:500; rabbit polyclonal antibody, bs-4564R, Bioss), cleaved caspase-3 (1:500; rabbit polyclonal antibody, WL01992, Wanleibio, Shenyang, China) and cleaved poly (ADP-ribose) polymerase (PARP) (1:500; rabbit polyclonal antibody, WL01932, Wanleibio), AKT (1:500; rabbit polyclonal antibody, bs-6951R, Bioss), or p-AKT (1:1,000; mouse monoclonal antibody, bsm-33281M, Bioss) were diluted in 5% skim milk.

Techniques: Western Blot

Journal: Cells

Article Title: A Novel Liquid Biopsy Method Based on Specific Combinations of Vesicular Markers Allows Us to Discriminate Prostate Cancer from Hyperplasia

doi: 10.3390/cells13151286

Figure Lengend Snippet: The cluster counts in the tumor and hyperplasia samples for the surface and intra-vesicular targets. The subfigures represent clusters expressing ERBB3 and ALK ( A ); ERBB3, ALK, and CD81 ( B ); STAT3 ( C ); STAT3 and CD81 ( D ); STAT3 and CyclinD1 ( E ); and STAT3, CyclinD1, and CD81 ( F ). Abbreviations: PCa: prostate cancer; BPH: benign prostate hyperplasia. Symbols represent: “*” p < 0.05; “ns” p ≥ 0.05; “•”refer to values out of ±1.5 * IQR.

Article Snippet: Additionally, custom antibodies, such as anti-ErbB-3/HER3-AF ® 555 (Bioss Antibodies, bs-1454R-A555), anti-ALK-AF ® 488 (Bioss Antibodies, bs-0097R-A488), anti-STAT3-AF ® 555 (Bioss Antibodies, bs-3429R-A555), and anti-Cyclin D1-AF ® 488 (Bioss Antibodies, bs-0623R-A488), were used.

Techniques: Expressing

Journal: Cells

Article Title: A Novel Liquid Biopsy Method Based on Specific Combinations of Vesicular Markers Allows Us to Discriminate Prostate Cancer from Hyperplasia

doi: 10.3390/cells13151286

Figure Lengend Snippet: The receiver operating characteristic curves for the intra-vesicular and surface targets. ROC curves for intra-vesicular and surface targets with the most differentiated counts: ERBB3, ALK ( A ); ERBB3, ALK, and CD81 ( B ); STAT3 ( C ); STAT3 and CD81 ( D ); STAT3 and CyclinD1 ( E ); and STAT3, CyclinD1, and CD81 ( F ).

Article Snippet: Additionally, custom antibodies, such as anti-ErbB-3/HER3-AF ® 555 (Bioss Antibodies, bs-1454R-A555), anti-ALK-AF ® 488 (Bioss Antibodies, bs-0097R-A488), anti-STAT3-AF ® 555 (Bioss Antibodies, bs-3429R-A555), and anti-Cyclin D1-AF ® 488 (Bioss Antibodies, bs-0623R-A488), were used.

Techniques:

Journal: Cells

Article Title: A Novel Liquid Biopsy Method Based on Specific Combinations of Vesicular Markers Allows Us to Discriminate Prostate Cancer from Hyperplasia

doi: 10.3390/cells13151286

Figure Lengend Snippet: Contingency tables with cutoff values and performance metrics for markers: ERBB3 and ALK; ERBB3, ALK, and CD81; STAT3; STAT3 and CD81; STAT3 and CyclinD1; and STAT3, CyclinD1, and CD81.

Article Snippet: Additionally, custom antibodies, such as anti-ErbB-3/HER3-AF ® 555 (Bioss Antibodies, bs-1454R-A555), anti-ALK-AF ® 488 (Bioss Antibodies, bs-0097R-A488), anti-STAT3-AF ® 555 (Bioss Antibodies, bs-3429R-A555), and anti-Cyclin D1-AF ® 488 (Bioss Antibodies, bs-0623R-A488), were used.

Techniques:

Journal: Cells

Article Title: A Novel Liquid Biopsy Method Based on Specific Combinations of Vesicular Markers Allows Us to Discriminate Prostate Cancer from Hyperplasia

doi: 10.3390/cells13151286

Figure Lengend Snippet: K-means cluster analysis. The above graph was generated from the analysis of the four most characterizing targets: STAT3; STAT3 and CD81; STAT3 and CyclinD1; and STAT3, CyclinD1, and CD81.

Article Snippet: Additionally, custom antibodies, such as anti-ErbB-3/HER3-AF ® 555 (Bioss Antibodies, bs-1454R-A555), anti-ALK-AF ® 488 (Bioss Antibodies, bs-0097R-A488), anti-STAT3-AF ® 555 (Bioss Antibodies, bs-3429R-A555), and anti-Cyclin D1-AF ® 488 (Bioss Antibodies, bs-0623R-A488), were used.

Techniques: Generated

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Gastric cancer-derived exosomal let-7 g-5p mediated by SERPINE1 promotes macrophage M2 polarization and gastric cancer progression

doi: 10.1186/s13046-024-03269-4

Figure Lengend Snippet: SERPINE1 -mediated GC-derived exosomal let-7 g-5p facilitates macrophage M2 polarization through STAT3 hyperphosphorylation resulting from inhibition of SOCS7 interactions with STAT3. ( A ) Differential miRNA analysis of exosomes derived from MKN45 cells with stably silenced SERPINE1 and normal MKN45 cells using sRNA-Seq. N, normal group. sh, stably silenced SERPINE1 . ( B ) Venn diagram of target genes predicted by miRDB, miRWalk, and miRTarBase for let-7 g-5p. ( C ) Network of target genes that interact with STAT3. ( D ) KEGG pathway analysis of the 78 target genes of let-7 g-5p using DAVID. ( E ) Schematic representation: exosomal let-7 g-5p ingested by macrophages inhibits SOCS7 interaction with STAT3, resulting in STAT3 hyperphosphorylation. ( F ) Flow cytometric assay of the impact of let-7 g-5p on M2 polarization induced by exosomes derived from GC cells. ( G ) Western blotting analysis for the levels of SOCS7 protein and STAT3 phosphorylation in macrophages treated with exosomes and antagomir-let-7 g-5p. ( H and I ) Endogenous CoIP assay for SOCS7 and STAT3 in macrophages ingesting exosomes derived from normal MKN45 cells. ( J ) Western blotting analysis of SOCS7 protein levels in xenograft tumors

Article Snippet: Antigen retrieval was performed using sodium citrate buffer (pH 6.0, 98 °C), followed by goat serum blocking for 1 h. Sections were incubated with PAI-1 (rabbit, 1:200, Immunoway), CD163 (mouse, 1:200, Immunoway), CD206 (mouse, 1:200, Proteintech), F4/80 (rabbit, 1:200, Bioss), iNOS (rabbit, 1:200, Bioss), Arg1 (rabbit, 1:200, Proteintech), STAT3 (rabbit, 1:200, Bioss) antibodies overnight at 4 °C, reactivated, stained with Cy3-conjugated goat anti-rabbit IgG (Abcam) and Alexa Fluor 488-conjugated goat anti-mouse IgG (Abcam) for 30 min, counterstained with DAPI, and imaged using fluorescence microscopy (IX51, Olympus) for ImageJ analysis.

Techniques: Derivative Assay, Inhibition, Stable Transfection, Flow Cytometry, Western Blot, Co-Immunoprecipitation Assay

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Gastric cancer-derived exosomal let-7 g-5p mediated by SERPINE1 promotes macrophage M2 polarization and gastric cancer progression

doi: 10.1186/s13046-024-03269-4

Figure Lengend Snippet: SERPINE1 promotes exosomal let-7 g-5p expression through the JAK2/STAT3 pathway. ( A ) GSEA was conducted for SERPINE1 co-expressed genes using GSEA (version 4.1.0). ( B ) Heatmap of 35 phosphorylation sites of 34 STAT3 upstream proteins determined by the median fluorescent intensity of the protein array normalized using Grubb’s algorithm. ( C ) Venn diagram of 34 downregulated phosphorylated proteins in the silenced SERPINE1 group and 107 TFs targeting let-7 g-5p. ( D ) Bubble plot combined with Sankey diagram of enriched KEGG pathways for 32 of the 34 STAT3 upstream proteins. ( E ) Statistical analysis of normalized phospho- and nonphospho-fluorescent protein spots in protein arrays. Western blotting analysis of total protein and phosphorylation levels of JAK2/STAT3 in GC cells silencing SERPINE1 ( F ), overexpressing SERPINE1 or treated with a JAK inhibitor ( G ). ( H ) Western blotting analysis of JAK2/STAT3 and SOCS7 in xenograft tumors. ( I ) Representative FISH images and comparison of let-7 g-5p expression in GC cells. ( J ) qRT-PCR analysis of exosomal let-7 g-5p expression in GC cells. ( K ) Representative FISH images and comparison of let-7 g-5p expression in xenograft tissues. ( L ) STAT3-binding motif and sites in the let-7 g-5p promoter region predicted using the JASPR database. ( M ) ChIP-qPCR assay demonstrated that STAT3 interacted with the let-7 g-5p promoter (site position: -1666~-1483). ( N ) Dual-luciferase reporter gene assay for the let-7 g-5p promoter region (position: -1692~-1420). ( O ) Luciferase activity of wt- and mut- let-7 g-5p promoter in the presence of vector or oeSTAT3. Mut, mutated-type. WT, wild-type. Vector, negative control plasmid. oeSTAT3, STAT3 overexpression plasmid

Article Snippet: Antigen retrieval was performed using sodium citrate buffer (pH 6.0, 98 °C), followed by goat serum blocking for 1 h. Sections were incubated with PAI-1 (rabbit, 1:200, Immunoway), CD163 (mouse, 1:200, Immunoway), CD206 (mouse, 1:200, Proteintech), F4/80 (rabbit, 1:200, Bioss), iNOS (rabbit, 1:200, Bioss), Arg1 (rabbit, 1:200, Proteintech), STAT3 (rabbit, 1:200, Bioss) antibodies overnight at 4 °C, reactivated, stained with Cy3-conjugated goat anti-rabbit IgG (Abcam) and Alexa Fluor 488-conjugated goat anti-mouse IgG (Abcam) for 30 min, counterstained with DAPI, and imaged using fluorescence microscopy (IX51, Olympus) for ImageJ analysis.

Techniques: Expressing, Protein Array, Western Blot, Comparison, Quantitative RT-PCR, Binding Assay, Luciferase, Reporter Gene Assay, Activity Assay, Plasmid Preparation, Negative Control, Over Expression