Journal: BMC cancer

Article Title: NR1D1 suppressed the growth of ovarian cancer by abrogating the JAK/STAT3 signaling pathway.

doi: 10.1186/s12885-021-08597-8

Figure Lengend Snippet: Fig. 2 NR1D1 induced cell cycle arrest and apoptosis in ovarian cancer cells. A Cell cycle of NR1D1 over-expressed cells was determined by flow cytometry. B The levels of cyclinD and cyclinE were determined by western blot. C Flow cytometry was performed to determine the cell cycle of NR1D1 silenced cells. D Western blot was performed to determine the levels of cyclinD and cyclinE. E Apoptosis of NR1D1 over-expressed cells was determined by flow cytometry. F After transfection with NR1D1 over-expression plasmid, the levels of activated caspase-3 and caspase-9 were determined. The results are presented as mean + SD

Article Snippet: Following blocking with 5% bovine serum albumin, the membranes were incubated with antibodies against NR1D1 (1:1000; Abclonal, Wuhan, China), cyclinD (1:1000; ABclonal), cyclinE (1:1000; Proteintech, Wuhan, China), SOCS3 (1:1000; ABclonal), JAK-1 (1:1000; Affinity, Changzhou, China), p-JAK1 (Tyr 1034/Tyr 1035; 1:1000; Affinity), JAK2 (1:500; Affinity), p-JAK2 (Tyr 1007/Tyr 1008, 1:1000; Affinity), STAT3 (1:500; Affinity), p-STAT3 (Tyr 705, 1:500; Affinity), β-actin (1:2000; Proteintech) at 4 °C overnight.

Techniques: Flow Cytometry, Western Blot, Transfection, Over Expression, Plasmid Preparation

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: PLoS ONE

Article Title: STAT6 knockdown using multiple siRNA sequences inhibits proliferation and induces apoptosis of human colorectal and breast cancer cell lines

doi: 10.1371/journal.pone.0207558

Figure Lengend Snippet: (A) Measure of STAT6 mRNA level in HT-29 cells 24 hours post-transfection. The graph represents the mean ± SEM of multiple independent experiments (n) obtained by real-time PCR. Results were analysed by ΔΔCt method for relative quantifications. The fold change is represented by the Y axis, and values are normalized to control cells. (B) STAT6 protein level analysis at 2, 5 and 7 days post-transfection in HT-29 cells. The graph represents the mean of the percentage of STAT6 positive cells ± SEM of multiple independent experiments (n) obtained by flow cytometry. (C) Measure of STAT6 mRNA level in ZR-75-1 cells 3 days post-transfection. The graph represents the mean ± SEM of multiple independent experiments (n) obtained by real-time PCR. Results were analysed by ΔΔCt method for relative quantifications. The fold change is represented by the Y axis, and values are normalized to control cells. (D) STAT6 protein level analysis after 4 and 7 days of transfection in ZR-75-1 cells. The graph represents the mean of the percentage of STAT6 positive cells ± SEM of multiple independent experiments (n) obtained by flow cytometry. E) Representative histograms (Control: back; NT: grey; STAT6.1: red; STAT6.4: blue) of STAT6 protein analysis at 7 days post-transfection by flow cytometry in HT-29 cells (top) and ZR-75-1 (bottom) cells. STAT6 siRNA sequences and non-targeting siRNA were used at 100 nM as the final concentration. Control cells were non-transfected cells and STAT6 siRNA sequences 1 and 4 and non-targeting siRNA are denoted as STAT6.1, STAT6.4 and NT, respectively. The number of independent experiments (n) is set out in the Fig.

Article Snippet: Cells were washed twice with PBS/0.5%BSA (Bovine Serum Albumin) and stained with anti-STAT6 APC conjugated antibody (Miltenyi Biotec, 130-104-030) (20 μl/10 6 cells) for 30 min in the dark at 4°C.

Techniques: Transfection, Real-time Polymerase Chain Reaction, Control, Flow Cytometry, Concentration Assay

Journal: PLoS ONE

Article Title: STAT6 knockdown using multiple siRNA sequences inhibits proliferation and induces apoptosis of human colorectal and breast cancer cell lines

doi: 10.1371/journal.pone.0207558

Figure Lengend Snippet: (A and B) Number of live HT-29 cells measured at 5 and 7 days post-transfection, respectively. The graphs represent the mean ± SEM of multiple independent experiments (n). (C) The graph illustrates how HT-29 cells grew over time and represents the mean ± SEM of the independent experiments (n) shown in A and B. (D and E) Number of live ZR-75-1 cells measured at 4 and 7 days post-transfection, respectively. The graphs represent the mean ± SEM of multiple independent experiments (n). (F) The graph illustrates how ZR-75-1 cells grew over time and represents the mean ± SEM of the multiple independent experiments (n) shown in D and E. The number of live cells was calculated as detailed in the material and methods using NucleoCounter NC-100. The percentage of reduction of the number of live cells was calculated by comparison between the mean of NT vs . the mean of the transfection with STAT6 siRNA sequences. STAT6 and non-targeting (NT) siRNA sequences were used at 100 nM as the final concentration. Non-transfected cells served as negative control and STAT6 siRNA sequences 1 and 4 and non-targeting siRNA are denoted as STAT6.1, STAT6.4 and NT, respectively. The number of independent experiments (n) is set out in the Fig.

Article Snippet: Cells were washed twice with PBS/0.5%BSA (Bovine Serum Albumin) and stained with anti-STAT6 APC conjugated antibody (Miltenyi Biotec, 130-104-030) (20 μl/10 6 cells) for 30 min in the dark at 4°C.

Techniques: Transfection, Comparison, Concentration Assay, Negative Control

Journal: PLoS ONE

Article Title: STAT6 knockdown using multiple siRNA sequences inhibits proliferation and induces apoptosis of human colorectal and breast cancer cell lines

doi: 10.1371/journal.pone.0207558

Figure Lengend Snippet: (A) Apoptosis analysis in HT-29 cells. (B) Apoptosis analysis in ZR-75-1 cells. In both cases, the graphs represent from left to right: early (Annexin V + /PI - ), late (Annexin V + /PI + ) and total (Annexin V + ) apoptosis. The graphs represent the mean ± SEM of multiple independent experiments (n) obtained by flow cytometry. (C) Representative flow cytometry plots in HT-29 cells. (D) Representative flow cytometry plots in ZR-75-1 cells. The X axes represent Annexin V and the Y axes represent PI fluorescence intensity. Quadrants were set according to cells independently stained with Annexin V or PI. Apoptosis was studied at 7 days post-transfection in both cell lines. Data were analysed with Flowjo Software. STAT6 siRNA sequences and a non-targeting siRNA sequence were used at 100 nM as the final concentration. Non-transfected cells served as control cells and STAT6 siRNA sequences 1 and 4 and non-targeting siRNA are denoted as STAT6.1, STAT6.4 and NT, respectively. The number of independent experiments (n) is set out in the Fig.

Article Snippet: Cells were washed twice with PBS/0.5%BSA (Bovine Serum Albumin) and stained with anti-STAT6 APC conjugated antibody (Miltenyi Biotec, 130-104-030) (20 μl/10 6 cells) for 30 min in the dark at 4°C.

Techniques: Flow Cytometry, Fluorescence, Staining, Transfection, Software, Sequencing, Concentration Assay, Control

Journal: PLoS ONE

Article Title: STAT6 knockdown using multiple siRNA sequences inhibits proliferation and induces apoptosis of human colorectal and breast cancer cell lines

doi: 10.1371/journal.pone.0207558

Figure Lengend Snippet: STAT6 siRNA transfection was carried out at day 1 of cell culture with (A) STAT6.1 and (B) STAT6.4 at 100 nM. A second transfection was carried out in both cases with STAT6.1 and STAT6.4 at the same concentration 7 days after the first transfection. The graphs represent the number of live cells over time measured at day 7 and 14 days post-first transfection counted using NucleoCounter NC-100 as detailed in the material and methods section. The mean ± SEM of 3 independent experiments is represented in each graph. Control cells were non-transfected cells and STAT6 siRNA sequences 1 and 4 and non-targeting siRNA are denoted as STAT6.1, STAT6.4 and NT, respectively. The percentage of reduction of the number of live cells was calculated by comparison between the mean of NT vs . the mean of the individual transfection with STAT6 siRNA sequences, and sequential transfection with NT (NT+NT) vs . sequential transfection with STAT6.1 and STAT6.4.

Article Snippet: Cells were washed twice with PBS/0.5%BSA (Bovine Serum Albumin) and stained with anti-STAT6 APC conjugated antibody (Miltenyi Biotec, 130-104-030) (20 μl/10 6 cells) for 30 min in the dark at 4°C.

Techniques: Transfection, Cell Culture, Concentration Assay, Control, Comparison

Journal: PLoS ONE

Article Title: STAT6 knockdown using multiple siRNA sequences inhibits proliferation and induces apoptosis of human colorectal and breast cancer cell lines

doi: 10.1371/journal.pone.0207558

Figure Lengend Snippet: (A) STAT6 expression at protein level in HT-29 cells at day 2 and 7 post-transfection measured by flow cytometry. The graph represents the mean ± SEM of multiple independent experiments (n). Data was analysed using Flowjo Software. The percentage of STAT6 positive cells is represented on the Y axis. (B) Representative histograms (Control: back; NT: grey; STAT6.1: red; STAT6.4: blue) of STAT6 protein analysis at 7 days post-transfection by flow cytometry in HT-29 cells. (C) Number of live HT-29 cells measured at day 7 post-transfection by NucleoCounter NC100. The graph represents the mean ± SEM of multiple independent experiments (n). (D) The graph illustrates how HT-29 cells grew over time and represents the mean ± SEM of the independent number of experiments shown in C. (E, F and G) The graphs show early (E) (Annexin V + /PI - ), late (F) (Annexin V + /PI + ) and total (G) (Annexin V + ) apoptosis, and represent the mean ± SEM of multiple independent experiments (n) obtained by flow cytometry. Apoptosis was studied at 7 days post-transfection. Data were analysed with Flowjo Software. STAT6 siRNA sequences and a non-targeting siRNA sequence were used at 100 nM as the final concentration. Non-transfected cells served as control cells and STAT6 siRNA sequences 1 and 4 and non-targeting siRNA are denoted as STAT6.1, STAT6.4 and NT, respectively. The number of independent experiments (n) is set out in the Fig.

Article Snippet: Cells were washed twice with PBS/0.5%BSA (Bovine Serum Albumin) and stained with anti-STAT6 APC conjugated antibody (Miltenyi Biotec, 130-104-030) (20 μl/10 6 cells) for 30 min in the dark at 4°C.

Techniques: Expressing, Transfection, Flow Cytometry, Software, Control, Sequencing, Concentration Assay

Journal: Turkish Journal of Biochemistry

Article Title: Effect of silibinin on GAS6/sAXL and JAK/STAT pathways in human cholangiocarcinoma cell line

doi: 10.1515/tjb-2022-0275

Figure Lengend Snippet: Figure 4: Silibinin significantly decreased the protein concentration of JAK2 (p<0.001). A. Silibinin significantly decreased the protein concentration of sAXL (p<0.001). B. Silibinin significantly increased the protein concentration of STAT1 p<0.001 C. Silibinin increased the protein concentration of GAS6 p>0.469 D. Error bars point out mean ± SD *, statistically significance at (*, p<0.01). All experiments were done at least in triplicate and compared to the control group.

Article Snippet: Assessment of JAK2, STAT1, GAS6 and sAXL protein concentrations JAK2, STAT1, GAS6, and sAXL protein concentrations were analyzed by ELISA according to the manufacturer’s instructions; JAK2, sAXL, and GAS6 ELISA kits were purchased from Bioassay technology laboratory (BT LAB, Shanghai Korain Biotech, Shanghai, China).

Techniques: Protein Concentration, Control