Journal: Asian Pacific Journal of Cancer Prevention : APJCP

Article Title: Possible Role of IL-6R/STAT3/MiRNA-34a Feedback Loop in Osteosarcoma

doi: 10.31557/APJCP.2023.24.9.3269

Figure Lengend Snippet: Box Plots for miR-34a, STAT3 and IL-6R Levels among Studied Groups. *, Significant difference versus control group; #, Significant difference versus non metastatic group; P value <0.05 is considered significant

Article Snippet: Estimation of STAT3 by ELISA technique STAT3 was measured using Human Signal Transducer and Activator of Transcription 3 ( STAT3 ) ELISA kit, BT LAB, Cat. No. E0650Hu according to the manufacturer’s instructions.

Techniques: Control

Journal: Asian Pacific Journal of Cancer Prevention : APJCP

Article Title: Possible Role of IL-6R/STAT3/MiRNA-34a Feedback Loop in Osteosarcoma

doi: 10.31557/APJCP.2023.24.9.3269

Figure Lengend Snippet: Level of miRNA-34a, STAT3 and IL-6R in the Studied Groups

Article Snippet: Estimation of STAT3 by ELISA technique STAT3 was measured using Human Signal Transducer and Activator of Transcription 3 ( STAT3 ) ELISA kit, BT LAB, Cat. No. E0650Hu according to the manufacturer’s instructions.

Techniques: Control, Expressing

Journal: Asian Pacific Journal of Cancer Prevention : APJCP

Article Title: Possible Role of IL-6R/STAT3/MiRNA-34a Feedback Loop in Osteosarcoma

doi: 10.31557/APJCP.2023.24.9.3269

Figure Lengend Snippet: (A) Significant positive correlation between STAT3 and IL6-R (r=0.868, p <0.001). (B) Significant inverse correlation between IL6 and miRNA-34a (r=-0.993, p <0.001)

Article Snippet: Estimation of STAT3 by ELISA technique STAT3 was measured using Human Signal Transducer and Activator of Transcription 3 ( STAT3 ) ELISA kit, BT LAB, Cat. No. E0650Hu according to the manufacturer’s instructions.

Techniques:

Journal: Asian Pacific Journal of Cancer Prevention : APJCP

Article Title: Possible Role of IL-6R/STAT3/MiRNA-34a Feedback Loop in Osteosarcoma

doi: 10.31557/APJCP.2023.24.9.3269

Figure Lengend Snippet: Correlation between Studied Parameters in the Non-Metastatic Group

Article Snippet: Estimation of STAT3 by ELISA technique STAT3 was measured using Human Signal Transducer and Activator of Transcription 3 ( STAT3 ) ELISA kit, BT LAB, Cat. No. E0650Hu according to the manufacturer’s instructions.

Techniques:

Journal: Asian Pacific Journal of Cancer Prevention : APJCP

Article Title: Possible Role of IL-6R/STAT3/MiRNA-34a Feedback Loop in Osteosarcoma

doi: 10.31557/APJCP.2023.24.9.3269

Figure Lengend Snippet: Correlation between Studied Parameters in the Metastatic Group

Article Snippet: Estimation of STAT3 by ELISA technique STAT3 was measured using Human Signal Transducer and Activator of Transcription 3 ( STAT3 ) ELISA kit, BT LAB, Cat. No. E0650Hu according to the manufacturer’s instructions.

Techniques:

Journal: Journal of Immunology Research

Article Title: TLR4-NLRP3-GSDMD-Mediated Pyroptosis Plays an Important Role in Aggravated Liver Injury of CD38 −/− Sepsis Mice

doi: 10.1155/2021/6687555

Figure Lengend Snippet: Escherichia coli can induce liver injury in mice. WT mice were injected with PBS or 3 × 10 8 cfu/ml E. coli intraperitoneally and sacrificed 3 hours later. Liver pathological injuries were observed with hematoxylin and eosin staining (a–d). The yellow arrows indicate edema, the blue arrows indicate inflammatory cell infiltration, the black arrows indicate punctate necrosis, and the green arrows indicate binucleate hepatocytes. Bacteria from the liver of PBS- or E. coli -stimulated WT mice were cultured in MH medium (e, f) overnight and identified by Gram staining (g). The serum AST (h) and ALT (i) concentrations were determined by using the detection kits. The mRNA of liver inflammatory cytokines TLR4 (j), NLRP3 (k), IL-1 β (l), and IL-18 (m) of PBS- or 3 × 10 8 cfu/ml E. coli -stimulated mice were measured by RT-qPCR. Data are presented as means ± standard deviation. Statistical significance was determined by the paired t -test ( n = 3, ∗ p < 0.05, ∗∗ p < 0.01).

Article Snippet: The primary antibodies anti-TLR4 rabbit mAb (1 : 500) (CST, USA), anti-TRIF rabbit mAb (1 : 1000) (Proteintech, USA), anti-MyD88 mouse mAb (1 : 2000) (Proteintech, USA), anti-NF- κ B p65 rabbit mAb (1 : 1000) (CST, USA), anti-phospho-NF- κ B p65 rabbit mAb (1 : 500) (CST, USA), anti-IL-6 mouse mAb (1 : 2000) (Proteintech, USA), anti-iNOS rabbit mAb (1 : 1000) (CST, USA), anti-BAX rabbit mAb (1 : 5000) (Proteintech, USA), anti-NLRP3 rabbit mAb (1 : 500) (Boster, China), anti-ASC rabbit mAb (1 : 500) (Affinity, China), anti-caspase-1 rabbit mAb (1 : 500) (Abcam, UK), anti-IL-1 β rabbit mAb (1 : 1000) (CST, USA), anti-IL-18 rabbit mAb (1 : 1000) (Affinity, China), anti-caspase-3 rabbit mAb (1 : 500) (CST, USA), anti-GSDMD rabbit mAb (1 : 500) (Affinity, China), anti-ERK1/2 rabbit mAb (1 : 1000) (CST, USA), anti-phospho-ERK1/2 rabbit mAb (1 : 2000) (CST, USA), and anti-GAPDH rabbit mAb (1 : 5000) (Proteintech, USA) were used.

Techniques: Injection, Staining, Bacteria, Cell Culture, Quantitative RT-PCR, Standard Deviation

Journal: Journal of Immunology Research

Article Title: TLR4-NLRP3-GSDMD-Mediated Pyroptosis Plays an Important Role in Aggravated Liver Injury of CD38 −/− Sepsis Mice

doi: 10.1155/2021/6687555

Figure Lengend Snippet: The expression levels of pyroptosis-related markers were detected by Western blot. The expressions of liver pyroptosis proteins in WT, CD38 −/− , and CD38 −/− TLR4 mut mice were detected at 3 hours after E. coli stimulation by Western blot. The expressions of NLRP3, ASC, procaspase-1, cleaved caspase-1, IL-1 β , IL-18, procaspase-3, and cleaved caspase-3 were measured (a). And relative levels of NLRP3 to GAPDH (b), ASC to GAPDH (c), cleaved to procaspase-1 (d), IL-1 β to GAPDH (e), IL-18 to GAPDH (f), and cleaved to procaspase-3 (g) were analyzed by ImageJ software. Data are presented as means ± standard deviation. Statistical significance was determined by one-way ANOVA ( n = 3, ∗ p < 0.05, ∗∗ p < 0.01).

Article Snippet: The primary antibodies anti-TLR4 rabbit mAb (1 : 500) (CST, USA), anti-TRIF rabbit mAb (1 : 1000) (Proteintech, USA), anti-MyD88 mouse mAb (1 : 2000) (Proteintech, USA), anti-NF- κ B p65 rabbit mAb (1 : 1000) (CST, USA), anti-phospho-NF- κ B p65 rabbit mAb (1 : 500) (CST, USA), anti-IL-6 mouse mAb (1 : 2000) (Proteintech, USA), anti-iNOS rabbit mAb (1 : 1000) (CST, USA), anti-BAX rabbit mAb (1 : 5000) (Proteintech, USA), anti-NLRP3 rabbit mAb (1 : 500) (Boster, China), anti-ASC rabbit mAb (1 : 500) (Affinity, China), anti-caspase-1 rabbit mAb (1 : 500) (Abcam, UK), anti-IL-1 β rabbit mAb (1 : 1000) (CST, USA), anti-IL-18 rabbit mAb (1 : 1000) (Affinity, China), anti-caspase-3 rabbit mAb (1 : 500) (CST, USA), anti-GSDMD rabbit mAb (1 : 500) (Affinity, China), anti-ERK1/2 rabbit mAb (1 : 1000) (CST, USA), anti-phospho-ERK1/2 rabbit mAb (1 : 2000) (CST, USA), and anti-GAPDH rabbit mAb (1 : 5000) (Proteintech, USA) were used.

Techniques: Expressing, Western Blot, Software, Standard Deviation

Journal: Oncotarget

Article Title: Structural composition of components of geoherb Moutan Cortex contributes to anti-diabetic nephropathy activity

doi: 10.18632/oncotarget.23771

Figure Lengend Snippet: Figure 1: Comparison on the down-regulation of Moutan Cortex from different regions on protein expression levels of ICAM-1,TGF-β1 and FN. HBZY-1 mesangial cells were treated with 200 μg/mL AGEs in the presence or absence of Moutan Cortex (MC) extract of 200 μg/mL. Aminoguanidine of 10 μM was used as the positive control while BSA (200 μg/mL) as blank control. (A) Western blotting was performed to compare the protein expression levels. (B–D) The grayscale scan results of ICAM-1, TGF-β1 and FN. a, Control; b, 200 μg/mL AGEs; c, Positive control aminoguanidine group; “d-m” represent MC from Anhui, Guizhou, Zhejiang, Henan, Hunan, Hebei, Sichuan, Chongqing, Shandong, Gansu. Data are expressed as means ± SD, n = 3. ###P < 0.001 vs. BSA group; *P < 0.05, **P < 0.01 and ***P < 0.001 vs. AGEs group; $P < 0.05, $$P < 0.01 and $$$P < 0.001 vs. MC from Anhui group.

Article Snippet: Rabbit anti-mouse FN, TGF-β1 and ICAM-1 monoclonal antibodies were offered by Boster Biological Engineering Co., Ltd. (Wuhan, China).

Techniques: Comparison, Expressing, Positive Control, Control, Western Blot

Journal: Oncotarget

Article Title: Structural composition of components of geoherb Moutan Cortex contributes to anti-diabetic nephropathy activity

doi: 10.18632/oncotarget.23771

Figure Lengend Snippet: Figure 2: Effect of Moutan Cortex from different regions on ICAM-1 and TGF-β1 protein expression levels in kidney of DN rats. After being treated with STZ and/or MC extract of 5g/kg or , immunohistochemistry was conducted to evaluate the expression levels of ICAM-1 (A) and TGF-β1 (B) of renal tissues. “a” represents normal control; “b” represents model group (DN rats); “c” represents Positive control 0.1 g/kg AG; “d-m” represent Gansu, Chongqing, Shangdong, Sichuan, Zhejiang, Anhui, Hunan, Guizhou, Hebei, Henan.

Article Snippet: Rabbit anti-mouse FN, TGF-β1 and ICAM-1 monoclonal antibodies were offered by Boster Biological Engineering Co., Ltd. (Wuhan, China).

Techniques: Expressing, Immunohistochemistry, Control, Positive Control

Journal: Cancer Cell International

Article Title: A novel chalcone derivative suppresses melanoma cell growth through targeting Fyn/Stat3 pathway

doi: 10.1186/s12935-020-01336-2

Figure Lengend Snippet: Lj-1-60 is a novel inhibitor targeting Fyn protein kinase. a HEK293T cells transfected with 2 μg and 4 μg of Fyn-flag plasmids for 36 h, it was then harvested and assessed by pull-down assay. Cell lysates incubated with Sepharose coupled with Lj-1-60 were subjected to SDS-PAGE, and analyzed by immunoblotting with antibody anti-flag. b Melanoma cell lysates for pull-down assay was detected by immunoblotting with anti-Fyn antibody. c In vitro kinase assay. A reaction mixture of substrate human recombinant protein Stat3 with myc-tag (676-770 aa), Fyn kinase and Lj-1-60 were incubated at 30 °C for 40 min and subjected to immunoblotting with indicated antibodies. Data are representative of three independent experiments. d Knocked down of Fyn in Sk-Mel-5 and Sk-Mel-28 cell lines by two independent shRNA. Total cell lysates were subjected to immunoblotting using indicated antibodies. Data are representative of three independent experiments. e Lj-1-60 treated in Sk-Mel-5 and Sk-Mel-28 cell lines for 48 h with indicated concentration and analyzed by immunoblot using antibodies p-Stat3(Tyr 705), Stat3, and GAPDH as loading control

Article Snippet: Recombinant human Stat3 was purchased from CUSABIO.

Techniques: Transfection, Pull Down Assay, Incubation, SDS Page, Western Blot, In Vitro, Kinase Assay, Recombinant, shRNA, Concentration Assay, Control

Journal: Cancer Cell International

Article Title: A novel chalcone derivative suppresses melanoma cell growth through targeting Fyn/Stat3 pathway

doi: 10.1186/s12935-020-01336-2

Figure Lengend Snippet: Analysis of gene expression profiles involved in melanoma cells altered by Lj-1-60. a The mRNA expression of CDKN1A, GADD45A, MCM3 in melanoma cells Sk-Mel-5 and Sk-Mel-28 was measured by RT-PCR. Total RNA was extracted from the cells treated with 2 μM Lj-1-60 for 24, 48 h. Data were expressed as mean (n = 3) ± SD, *P < 0.05, ***P < 0.001, ****P < 0.0001. b The mRNA expression of CDKN1A, GADD45A, MCM3 in melanoma cells Sk-Mel-5 and Sk-Mel-28 was examined by RT-PCR. Total RNA was extracted from cells knocked down of Fyn with two independent shRNA. Data were expressed as mean (n = 3) ± SD, *P < 0.05, ***P < 0.001, ****P < 0.0001. c Structural model. Lj-1-60 inhibits the proliferation of melanoma and induces cell cycle arrested in G2/M phase and apoptosis by targeting Fyn through inhibiting the phosphorylation of Stat3

Article Snippet: Recombinant human Stat3 was purchased from CUSABIO.

Techniques: Gene Expression, Expressing, Reverse Transcription Polymerase Chain Reaction, shRNA, Phospho-proteomics

Journal: OncoTargets and Therapy

Article Title:

Knockdown of ROS proto-oncogene 1 inhibits migration and invasion in gastric cancer cells by targeting the PI3K/Akt signaling pathway

doi: 10.2147/ott.s213421

Figure Lengend Snippet: Figure 7 The levels of PI3K, p-PI3K, Akt and p-Akt were quantified by Western blotting. The ratios of p-PI3K/PI3K and p-Akt/Akt were calculated. The results were shown as mean±SD. **P<0.01 versus the shCtr group.

Article Snippet: After blocking, the membranes were incubated with primary antibodies against ROS1 (1:500, Sangon Biotech, Shanghai, China), cleaved-caspase-3 (1:1000, Abcam, Cambridge Science Park, Cambridge, UK), Bcl-2 (1:400, BOSTER, Wuhan, China), Bax (1:400, BOSTER), cleaved-PARP (1:1000, Abcam), E-cadherin (1:400, BOSTER), Vimentin (1:500, BIOSS, Beijing, China), Ncadherin (1:400, BOSTER), p-PI3K (1:500, BIOSS), PI3K (1:400, BOSTER), p-Akt (1:200, Santa Cruz Biotechnology, Dallas, Texas, USA) and Akt (1:200, Santa Cruz Biotechnology) at 4°C overnight.

Techniques: Western Blot

Journal: The FASEB Journal

Article Title: Activation of JAK/STAT3 restores NK-cell function and improves immune defense after brain ischemia

doi: 10.1096/fj.201700962R

Figure Lengend Snippet: Gene overlap between poststroke NK-cell gene profile and NK suppression–induced infection gene network

Article Snippet: CRISPR activation plasmid cDNA transfection STAT3 clustered regularly interspaced short palindromic repeats (CRISPR) activation plasmid (sc-423176-ACT; Santa Cruz Biotechnology, Dallas, TX, USA) was used to activate and overexpress STAT3 in NK cells.

Techniques: Infection, Virus

Journal: The FASEB Journal

Article Title: Activation of JAK/STAT3 restores NK-cell function and improves immune defense after brain ischemia

doi: 10.1096/fj.201700962R

Figure Lengend Snippet: Genetic activation of STAT3 preserved NK-cell–derived IFN-γ expression after MCAO. C57BL/6 mice were subjected to sham operation or MCAO surgery. A, B) Real-time PCR and flow cytometry plots showed STAT3 gene (A) and protein levels (B) in splenic NK cells 24 h after MCAO; n = 4 per group. C) The schematic graph of the experimental design. Splenic NK cells isolated from wild-type mice were treated with STAT3-CRISPR activation plasmid or control vector. After transfected for 48 h, 3 × 106 NK cells were transferred intraveniously into groups of Rag2−/−γc−/− mice before MCAO. D) Flow cytometry plots and bar graph showed STAT3 phosphorylation in NK cells 2 h after MCAO; n = 4/group. E, F) Flow cytometry plots (E) and bar graph (F) show the effect of STAT3 activation on functional marker expression (CD69, perforin, and IFN-γ) of splenic NK cells at d 1 after MCAO. Plots represent the results from 3 independent experiments. Error bars represent mean ± sem. **P < 0.01 by 2-tailed, unpaired Student’s t test.

Article Snippet: CRISPR activation plasmid cDNA transfection STAT3 clustered regularly interspaced short palindromic repeats (CRISPR) activation plasmid (sc-423176-ACT; Santa Cruz Biotechnology, Dallas, TX, USA) was used to activate and overexpress STAT3 in NK cells.

Techniques: Activation Assay, Derivative Assay, Expressing, Real-time Polymerase Chain Reaction, Flow Cytometry, Isolation, CRISPR, Plasmid Preparation, Control, Transfection, Phospho-proteomics, Functional Assay, Marker

Journal: The FASEB Journal

Article Title: Activation of JAK/STAT3 restores NK-cell function and improves immune defense after brain ischemia

doi: 10.1096/fj.201700962R

Figure Lengend Snippet: STAT3 activation in NK cells improves survival and reduces lung bacterial burden in MCAO mice. Splenic NK cells isolated from C57BL/6 mice targeting STAT3 were established by introducing a STAT3 activation plasmid. STAT3-activated (STAT3-CRISPR) NK cells were then passively transferred via intravenious injection to Rag2−/−γc−/− transgenic mice, followed by MCAO surgery. A) Representative MRI images and summarized data showed stroke lesion volume after adoptive transfer with STAT3-CRISPR NK cells; n = 5 mice/group. B) The neurodeficit score was assessed by modified neurological severity score (mNSS) after STAT3-CRISPR NK-cell transfer and MCAO surgery. P > 0.05 by 2-way ANOVA (A, B). C) The survival of the mice was noted at 0 to 8 d after cell transfer and MCAO surgery; n = 15 mice/group. *P < 0.05 by 2-way ANOVA. D) Lung tissues from MCAO mice transferred with STAT3-CRISPR NK cells were collected for bacteriologic analysis at 3 d after MCAO surgery. Data summarized in D graphically illustrate the quantification of bacteria burden in lung after passive transfer with NK cells. Data are presented in colony-forming units (CFU) per organ lung tissue homogenate. **P < 0.01 by 2-tailed, unpaired Student’s t test. E) Hematoxylin and eosin–stained lung sections exhibited typical signs (thickening of alveolar walls and neutrophilic infiltrates) of bacterial burden in MCAO mice. The lung tissues from wild-type mice adoptively transferred with control vector transfected-NK or STAT3-CRISPR NK cells were collected at d 3 after MCAO for histologic examination. Scale bars, 50 μm. F) ELISA measurement of IFN-γ protein levels in serum from the control or STAT3-CRISPR NK groups at d 3 after MCAO. **P < 0.01 by 2-tailed, unpaired Student’s t test. Mean ± sem.

Article Snippet: CRISPR activation plasmid cDNA transfection STAT3 clustered regularly interspaced short palindromic repeats (CRISPR) activation plasmid (sc-423176-ACT; Santa Cruz Biotechnology, Dallas, TX, USA) was used to activate and overexpress STAT3 in NK cells.

Techniques: Activation Assay, Isolation, Plasmid Preparation, CRISPR, Injection, Transgenic Assay, Adoptive Transfer Assay, Modification, Bacteria, Staining, Control, Transfection, Enzyme-linked Immunosorbent Assay