Journal: Infection and Immunity

Article Title: STAT3 Promotes Schistosome-Induced Liver Injury by Inflammation, Oxidative Stress, Proliferation, and Apoptosis Signal Pathway

doi: 10.1128/iai.00309-20

Figure Lengend Snippet: Figure 3 JAK2/STAT3 signal pathway was activated in the liver of mice with S. 623

Article Snippet: After dissolution in 10% or 12% SDS-PAGE, 433 the total protein samples were transferred to nitrocellulose membranes for 2 h with 434 transfer buffer and blocked with 5% nonfat dried milk, which were dissolved in TBST 435 (20 mmol/L Tris–HCl, pH 7.6, 150 mmol/L NaCl and 0.02% Tween 20) for 1 h. 436 Membranes were incubated with primary antibodies for GAPDH (Cell Signaling 437 Technology-2118, 1:1500 dilution), α-SMA (abcam-ab5694,1:800), IL-6 438 (abcam-ab6672), p-JAK2 (Cell Signaling Technology-3776), JAK2 (Cell Signaling 439 Technology-3230), p-STAT3 (Cell Signaling Technology-9145), STAT3 (Cell 440 on D ecem ber 17, 2020 at 43709427 http://iai.asm .org/ D ow nloaded from 21 Signaling Technology-9139/4904), Bax (Santa Cruz Biotechnology-SC493), Bcl-2 441 (Santa Cruz Biotechnology-SC492), Sirt3 (Cell Signaling Technology-5490s), 442 Acetylated SOD2 (K68) (abcam-ab137037), SOD2 (abcam-ab13534) overnight at 443 4°C.

Techniques:

Journal: Infection and Immunity

Article Title: STAT3 Promotes Schistosome-Induced Liver Injury by Inflammation, Oxidative Stress, Proliferation, and Apoptosis Signal Pathway

doi: 10.1128/iai.00309-20

Figure Lengend Snippet: Figure 4 Liver p-STAT3 deficiency attenuates hepatic injury caused by S. 635

Article Snippet: After dissolution in 10% or 12% SDS-PAGE, 433 the total protein samples were transferred to nitrocellulose membranes for 2 h with 434 transfer buffer and blocked with 5% nonfat dried milk, which were dissolved in TBST 435 (20 mmol/L Tris–HCl, pH 7.6, 150 mmol/L NaCl and 0.02% Tween 20) for 1 h. 436 Membranes were incubated with primary antibodies for GAPDH (Cell Signaling 437 Technology-2118, 1:1500 dilution), α-SMA (abcam-ab5694,1:800), IL-6 438 (abcam-ab6672), p-JAK2 (Cell Signaling Technology-3776), JAK2 (Cell Signaling 439 Technology-3230), p-STAT3 (Cell Signaling Technology-9145), STAT3 (Cell 440 on D ecem ber 17, 2020 at 43709427 http://iai.asm .org/ D ow nloaded from 21 Signaling Technology-9139/4904), Bax (Santa Cruz Biotechnology-SC493), Bcl-2 441 (Santa Cruz Biotechnology-SC492), Sirt3 (Cell Signaling Technology-5490s), 442 Acetylated SOD2 (K68) (abcam-ab137037), SOD2 (abcam-ab13534) overnight at 443 4°C.

Techniques:

Journal: Infection and Immunity

Article Title: STAT3 Promotes Schistosome-Induced Liver Injury by Inflammation, Oxidative Stress, Proliferation, and Apoptosis Signal Pathway

doi: 10.1128/iai.00309-20

Figure Lengend Snippet: Figure 6 Schematic overview of JAK2/STAT3-mediated effects on S. japonicum 668

Article Snippet: After dissolution in 10% or 12% SDS-PAGE, 433 the total protein samples were transferred to nitrocellulose membranes for 2 h with 434 transfer buffer and blocked with 5% nonfat dried milk, which were dissolved in TBST 435 (20 mmol/L Tris–HCl, pH 7.6, 150 mmol/L NaCl and 0.02% Tween 20) for 1 h. 436 Membranes were incubated with primary antibodies for GAPDH (Cell Signaling 437 Technology-2118, 1:1500 dilution), α-SMA (abcam-ab5694,1:800), IL-6 438 (abcam-ab6672), p-JAK2 (Cell Signaling Technology-3776), JAK2 (Cell Signaling 439 Technology-3230), p-STAT3 (Cell Signaling Technology-9145), STAT3 (Cell 440 on D ecem ber 17, 2020 at 43709427 http://iai.asm .org/ D ow nloaded from 21 Signaling Technology-9139/4904), Bax (Santa Cruz Biotechnology-SC493), Bcl-2 441 (Santa Cruz Biotechnology-SC492), Sirt3 (Cell Signaling Technology-5490s), 442 Acetylated SOD2 (K68) (abcam-ab137037), SOD2 (abcam-ab13534) overnight at 443 4°C.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Nimodipine Used with Vincristine: Protects Schwann Cells and Neuronal Cells from Vincristine-Induced Cell Death but Increases Tumor Cell Susceptibility

doi: 10.3390/ijms251910389

Figure Lengend Snippet: List of used antibodies.

Article Snippet: STAT3 (124H6) #9139 , Mouse IgG2a , 1:1000 , 5% MP in TBS-T , Cell Signaling Technology (Danvers, MA, USA).

Techniques:

Journal: bioRxiv

Article Title: Activated Oncostatin M signaling drives cancer-associated skeletal muscle wasting

doi: 10.1101/2023.01.26.525658

Figure Lengend Snippet: (A) Mouse primary myotubes were treated with recombinant OSM, IL6 or LIF (each 250 ng/ml) for 10 min. Protein levels were determined by western blotting. (B) Mouse primary myotubes were treated with Ruxolitinib (Rux; 2 μM) for 30 min and then recombinant OSM (250 ng/ml) was added for 10 min. Protein levels were determined by western blotting. (C) Mouse primary myotubes were treated with Ruxolitinib (Rux; 2 μM) and recombinant OSM (250 ng/ml) for 48 hr. mRNA levels were determined by RT-qPCR (n = 3 for each group). * p < 0.05, ** p < 0.01, *** p < 0.001 compares differences between Control and OSM groups. ## p < 0.01, ### p < 0.001 compares differences between OSM and Rux + OSM groups. (D-G) Mouse primary myotubes were transduced with LacZ or dominant-negative (dn) Stat3-Y705F expressing adenoviruses and treated with recombinant OSM (250 ng/ml) for 48 hr. Protein levels were determined by western blotting (D). mRNA levels were tested by RT-qPCR (n = 3 for each group) (E). Cells were also transduced with a GFP adenovirus for fluorescence imaging. Average myotube diameter was measured (n = 4 for each group) (F). Myotubes were visualized under the fluorescence microscope. Scale bar is 50 μm (G). * p < 0.05, ** p < 0.01, *** p < 0.001 compares differences between Ad-LacZ and Ad-LacZ + OSM groups. # p < 0.05, ## p < 0.01, ### p < 0.001 compares differences between Ad-LacZ + OSM and Ad-Stat3-Y705F + OSM groups. Data are presented as mean ± SEM. Statistical analysis was conducted using one-way ANOVA with Tukey’s post-hoc test.

Article Snippet: After blocking with 5% non-fat milk in Tris-buffered saline containing 0.05% Tween 20 (TBS-T), the membrane was blotted overnight with primary antibodies in TBS-T containing 5% BSA (Cell signaling antibodies; Stat1 (9172), phospho-Stat1-Tyr701-(7649), Stat3 (9139), phospho-Stat3-Tyr705 (9145), Stat5 (94205T), phospho-Stat5-Tyr694 (4322T), Jak2 (3230), phospho-Jak2-Tyr1008 (8082), β-actin (3700), p65-RelA (8242), phospho-p65-RelA-Ser536 (3033), p105/p50-NFκB (12540), phospho-p105-NFκB-Ser932 (4806), p100/p52-NFκB2 (4882), IκB (4814), phospho-IκB-Ser32 (2859), ECM Bioscience antibodies; Atrogin1 (AP2041) and MURF1 (MP3401), and DSHB; MyHC antibody (MF20)).

Techniques: Recombinant, Western Blot, Quantitative RT-PCR, Control, Transduction, Dominant Negative Mutation, Expressing, Fluorescence, Imaging, Microscopy

Journal: The Journal of Biological Chemistry

Article Title: Simultaneous isolation of hormone receptor–positive breast cancer organoids and fibroblasts reveals stroma-mediated resistance mechanisms

doi: 10.1016/j.jbc.2023.105021

Figure Lengend Snippet: CAF-secreted cytokines drive resistance to fulvestrant. A , quantification of the signal intensity of the five highly secreted cytokines from the cytokine arrays of CAF-CM (patients #1-27) normalized to cancer cell-CM. B , cell viability of MCF7 cells treated with recombinant human GROα/CXCL1, IL-8, CXCL5, HGF, CCL19, or PBS (100 ng/ml) and fulvestrant (500 nM). Presto Blue was used to assess cell viability after 72 h. Data are normalized to PBS treated groups and is a representative of three independent experiments. Error bars are SEM. C and D , quantification of EdU+ cells in PDOs treated with a cytokine cocktail and fulvestrant (500 nM) ( C ) or with recombinant human GROα, IL-8, CXCL5, HGF, CCL19, or PBS (100 ng/ml) and fulvestrant (500 nM) from two independent experiments. Proliferation was assessed by pulsing PDOs with EdU for 4 h and quantifying the ratio of EdU+ cells per total (DAPI+) number of cells in 20 PDOs. Red line indicates the mean. E , representative confocal images of PDOs treated with recombinant human GROα, HGF, CCL19, or PBS (100 ng/ml) and fulvestrant (500 nM). F , Western blot analysis of CAF-CM stimulated signaling pathway proteins pSTAT3, STAT3, ER, pAKT, AKT, pERK1/2, and ERK1/2 in PDOs grown in NC or CAF-CM and treated with 500 nM fulvestrant for 96 h. G , Western blot analysis of GROα stimulated signaling pathway proteins pSTAT3, STAT3, pERK1/2, and ERK1/2 in MCF7 cells starved overnight and incubated for 30 min with 100 ng/ml GROα. VCL was used as loading control. Student’s t test was used to determine significance by comparing each treatment to the PBS control in ( B ) and ( D ). The scale bar represents 40 μm. CAF, cancer-associated fibroblasts; DAPI, 4′,6-diamidino-2-phenylindole; EdU, 5-ethynyl-2′-deoxyuridine; ER, estrogen receptor; GROα, growth-regulated oncogene α; HGF, hepatocyte growth factor; IL, interleukins; PDOs, patient-derived organoids; VCL, vinculin.

Article Snippet: The membranes were blocked with 5% nonfat milk in 0.1% Tween 20 in Tris-buffered saline for 1 h and incubated with primary antibodies for 1 h in RT or overnight at 4 °C using the following antibodies: FN1 (Thermo Fisher Scientific #MA5-11981), PDGFRα (CST #3174), VIM (CST #5741), αSMA (Abcam #5694), CAV1 (CST #3267), FAP (CST #66562), PDPN (CST #9047), THY1 (CST #13801), pSTAT3 Tyr705 (CST#9145), STAT3 (CST#9139), ER (CST #13258), pAKT Ser473 (CST#4060), AKT (CST #9272), pERK1/2 Thr202/Tyr204 (CST #4370), ERK1/2 (CST #4695) and VCL (CST #13901).

Techniques: Recombinant, Western Blot, Incubation, Control, Derivative Assay