Journal: The Journal of Neuroscience

Article Title: Prolactin Regulates Tuberoinfundibular Dopamine Neuron Discharge Pattern: Novel Feedback Control Mechanisms in the Lactotrophic Axis

doi: 10.1523/JNEUROSCI.0129-12.2012

Figure Lengend Snippet: The LV and HV components of IPrl are activated by distinct signal transduction pathways. A, I–V relationship of IPrl in control (green; n = 14), wortmannin (200 nm; red; n = 8), and AG490 (10 μm; blue; n = 3). Wortmannin, but not AG490, significantly modifies IPrl at VHold = +40 mV. Values expressed as mean ± SEM. ***p < 0.005. B, I–V relationship of IPrl at a concentration of 500 nm Prl (green; n = 14), 40 nm Prl in which both the LV and HV components were present (red; n = 13 of 19), and 40 nm Prl in which only the LV component was present (blue; n = 6 of 19). Values expressed as mean ± SEM. ***p < 0.005.

Article Snippet: TTX was purchased from Alomone Labs. AG490 [( E )-2-cyano-3-(3,4-dihydrophenyl)- N -(phenylmethyl)-2-propenamide], nimodipine, paxilline, {"type":"entrez-protein","attrs":{"text":"SKF96365","term_id":"1156357400","term_text":"SKF96365"}} SKF96365 (1-[2-(4-methoxyphenyl)-2-[3-(4-methoxyphenyl)propoxy]ethyl-1 H -imidazole hydrochloride), UCL1648 [6,10-diaza-3(1,3)8,(1,4)-dibenzena-1,5(1,4)-diquinolinacy clodecaphane], and wortmannin were purchased from Tocris Bioscience. fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window Figure 6. caption a7 Prl broadens TIDA APs.

Techniques: Transduction, Control, Concentration Assay

Journal: Frontiers in physiology

Article Title: Acute exercise reduces feeding by activating IL-6/Tubby axis in the mouse hypothalamus.

doi: 10.3389/fphys.2022.956116

Figure Lengend Snippet: FIGURE 3 Hypothalamic IL-6 action through JAK2 to activate TUB. (A) Evaluation of cumulative food intake (g) at 4 h, 12 h, and 24 h (n = 5 in each group); Western blot showing (B) TUB tyrosine phosphorylation and (C) TUB associated with JAK2 from hypothalamic lysates from wt/wt mice (6–8 weeks old) obtained in the colony of B6-tub/tub mice under resting conditions (Sed). The mice were treated with vehicle or AG490 (JAK2inhibitor), IL-6, or IL-6 with AG490 by intracerebroventricular (ICV) injections. We used male mice. All values are expressed as means ± standard deviation (SD). Western blot assays (n = 4 in each group). Two-way ANOVA was used to analyze A, while one-way ANOVA with Tukey’s multiple comparisons tests was used to analyze B and C. *ICV IL-6 vs. Veh, AG490, and IL-6 plus AG490 (p < 0.0078); #ICV IL-6 vs. Veh, AG490 and IL-6 plus AG490 (p < 0.0041); ϕ ICV IL-6 vs. Veh, AG490, and IL-6 plus AG490 (p < 0.0136).

Article Snippet: ICV injection of AG490 (50 mM) (cat #0414, Tocris Bioscience, Bristol, United Kingdom) was used to inhibit JAK2 as previously described (Damm et al., 2013; Quaresma et al., 2017).

Techniques: Western Blot, Phospho-proteomics, Standard Deviation

Journal: International Journal of Biological Sciences

Article Title: An Autocrine IL-6/IGF-1R Loop Mediates EMT and Promotes Tumor Growth in Non-small Cell Lung Cancer

doi: 10.7150/ijbs.31999

Figure Lengend Snippet: IL-6 induced EMT through activation of STAT3. Cells were transfected with either STAT3 siRNA (siSTAT3) or negative control siRNA (NC siRNA) and activation of STAT3 was measured by western blot (A) . Cells were incubated with IL-6 (50ng/ml) with or without AG 490 (50uM/L) for 48 hours and harvested for western blot of EMT markers (B) and JAK/STAT3 signaling molecules (D) . Cells were transfected with siSTAT3 and EMT markers (C) and JAK/STAT3 signaling molecules (E) were measured by western blotting. Cells were treated with IL-6 with or without IL-6 antibody and EMT markers were measured (F) . Blots were representatives of three experiments.

Article Snippet: The inhibitors, dissolved in dimethyl sulfoxide (DMSO), were used in the following concentrations: STAT3 inhibitor AG-490 (50uM/L) (Selleck Chemicals, Houston, USA), IGF-1R inhibitor AG-1024 (5uM/L) (Selleck Chemicals), AKT inhibitor MK-2206 2HCl (5uM/L) (Selleck Chemicals, Houston, USA), JAK1 inhibitor ZM 39923 HCl (50uM /L) (Selleck Chemicals, Houston, USA).

Techniques: Activation Assay, Transfection, Negative Control, Western Blot, Incubation

Journal: International Journal of Biological Sciences

Article Title: An Autocrine IL-6/IGF-1R Loop Mediates EMT and Promotes Tumor Growth in Non-small Cell Lung Cancer

doi: 10.7150/ijbs.31999

Figure Lengend Snippet: IL-6 and IGF-1R cross-signal to promote EMT. PC-9 cells were treated with IL-6 with or without IGF-1R inhibitor AG1024 for 48 hours, and IGF-1R pathway (A) and JAK/STAT3 pathway (B) were tested using western blot. Cells were treated with IL-6 with or without STAT3 inhibitor AG490 for 48 hours, and IGF-1R pathway were tested using western blot (C) . STAT3 over-expressed plasmids were constructed and promoter activity of IGF-1R was studied by reporter assay (D) . Blots were representatives of three experiments and results were quantified using Quantity One for comparisons between IL-6 group and control or between IL-6 plus an inhibitor co-treated cells and IL-6 group. *, p<0.05. #, p>0.05 not significant.

Article Snippet: The inhibitors, dissolved in dimethyl sulfoxide (DMSO), were used in the following concentrations: STAT3 inhibitor AG-490 (50uM/L) (Selleck Chemicals, Houston, USA), IGF-1R inhibitor AG-1024 (5uM/L) (Selleck Chemicals), AKT inhibitor MK-2206 2HCl (5uM/L) (Selleck Chemicals, Houston, USA), JAK1 inhibitor ZM 39923 HCl (50uM /L) (Selleck Chemicals, Houston, USA).

Techniques: Western Blot, Construct, Activity Assay, Reporter Assay, Control

Journal: International Journal of Biological Sciences

Article Title: An Autocrine IL-6/IGF-1R Loop Mediates EMT and Promotes Tumor Growth in Non-small Cell Lung Cancer

doi: 10.7150/ijbs.31999

Figure Lengend Snippet: IL-6/ IGF-1R autocrine loop is mediated via STAT3. Cells were treated with IL-6 with or without an inhibitor (AG490, AG1024 or AKT inhibitor MK-2206 2HCl) and RT-PCR analyses were performed after 48 hours. Gene expressions of IL-6 (A and E) , IGF-1 (B and F) and IGF-2 (C and G) were compared between IL-6-treated cells and control group and between IL-6 plus inhibitor co-treated cells and IL-6 group. ELISAs were performed for IGF-1 (D) and IL-6 (H) and results were compared in the same way as the above mentioned RT-PCR analyses. *, p<0.05. **, p<0.01. #, p>0.05 not significant.

Article Snippet: The inhibitors, dissolved in dimethyl sulfoxide (DMSO), were used in the following concentrations: STAT3 inhibitor AG-490 (50uM/L) (Selleck Chemicals, Houston, USA), IGF-1R inhibitor AG-1024 (5uM/L) (Selleck Chemicals), AKT inhibitor MK-2206 2HCl (5uM/L) (Selleck Chemicals, Houston, USA), JAK1 inhibitor ZM 39923 HCl (50uM /L) (Selleck Chemicals, Houston, USA).

Techniques: Reverse Transcription Polymerase Chain Reaction, Control

Journal: International Journal of Biological Sciences

Article Title: An Autocrine IL-6/IGF-1R Loop Mediates EMT and Promotes Tumor Growth in Non-small Cell Lung Cancer

doi: 10.7150/ijbs.31999

Figure Lengend Snippet: Inhibiting STAT3 partly restores gefitinib sensitivity. (A to D) PC-9 cells were treated with gefitinib (50nmol/L) for 48 h. Cells were stained with annexin V-FITC and PI and subsequently analyzed using flow cytometry. (E) Quantification of gefitinib-induced apoptotic cells by annexin/PI double-staining. Apoptosis was evaluated by determining the percentage of annexin V-positive cells. The data are representative of three independent experiments and compared between IL-6 treated cells and gefitinib group, or between IL-6 plus an inhibitor co-treated cells and IL-6 group. *, p<0.05. **, p<0.01. #, p>0.05 not significant.

Article Snippet: The inhibitors, dissolved in dimethyl sulfoxide (DMSO), were used in the following concentrations: STAT3 inhibitor AG-490 (50uM/L) (Selleck Chemicals, Houston, USA), IGF-1R inhibitor AG-1024 (5uM/L) (Selleck Chemicals), AKT inhibitor MK-2206 2HCl (5uM/L) (Selleck Chemicals, Houston, USA), JAK1 inhibitor ZM 39923 HCl (50uM /L) (Selleck Chemicals, Houston, USA).

Techniques: Staining, Flow Cytometry, Double Staining

Journal: International Journal of Biological Sciences

Article Title: An Autocrine IL-6/IGF-1R Loop Mediates EMT and Promotes Tumor Growth in Non-small Cell Lung Cancer

doi: 10.7150/ijbs.31999

Figure Lengend Snippet: Inhibiting either STAT3 or IGF-1R attenuates cell migration. PC-9 cells (A) and A549 cells (B) were seeded and treated either with IL-6 alone or in the presence of both IL-6 and an STAT3 or IGF-1R inhibitor. Migration was shown as photograph and the number of migratory cells was determined using a transwell matrix penetration assay and quantified for analysis of significance. *, p<0.05. #, p>0.05 not significant.

Article Snippet: The inhibitors, dissolved in dimethyl sulfoxide (DMSO), were used in the following concentrations: STAT3 inhibitor AG-490 (50uM/L) (Selleck Chemicals, Houston, USA), IGF-1R inhibitor AG-1024 (5uM/L) (Selleck Chemicals), AKT inhibitor MK-2206 2HCl (5uM/L) (Selleck Chemicals, Houston, USA), JAK1 inhibitor ZM 39923 HCl (50uM /L) (Selleck Chemicals, Houston, USA).

Techniques: Migration

Journal: Journal of molecular and cellular cardiology

Article Title: Differential hypertrophic effects of cardiotrophin-1 on adult cardiomyocytes from normotensive and spontaneously hypertensive rats.

doi: 10.1016/j.yjmcc.2006.03.433

Figure Lengend Snippet: Fig. 5. Cells were pre-incubated with the indicated chemical inhibitors for 1 h and then with CT-1 for 48 h. Cell area (A), length (B) and width (C) were determined by planimetry. Values represent the mean±standard error of a minimum of 120 cardiomyocytes per experimental condition. Three independent experiments performed in three rats yielded identical results. **p<0.01 vs. baseline; *p<0.05 vs. baseline; $$p<0.01 vs. CT-1; $p<0.05 vs. CT-1. W, Wortmannin; PD1, PD98059 at 10−6 mol/L to inhibit p42/44; PD30, PD98059 at 30×10−6 mol/L to inhibit ERK-5; AG, AG490.

Article Snippet: 10− 6 mol/L to inhibit ERK5 (Calbiochem), the STATs inhibitor AG490 at 10−6 mol/L (Calbiochem), antibodies against LIFR (Santa Cruz) and gp130 (Santa Cruz) at 10−6 mol/L.

Techniques: Incubation

Journal: Journal of molecular and cellular cardiology

Article Title: Differential hypertrophic effects of cardiotrophin-1 on adult cardiomyocytes from normotensive and spontaneously hypertensive rats.

doi: 10.1016/j.yjmcc.2006.03.433

Figure Lengend Snippet: Fig. 6. (A) SHR cardiomyocytes were pre-incubated for 1 h with the STAT3 blocker AG490 and then with CT-1 for 24 h. A representative Western blot and the histogram with bars representing the mean±standard error of three independent experiments are shown for each protein. *p<0.01 vs. baseline and CT-1+AG. AG, AG490 (B) Angiotensinogen mRNA was assayed in cardiomyocytes from Wistar rats and SHR before and after 3 h incubation with CT-1. Bars represent mean±standard error of triplicates in three independent experiments. $p<0.01 vs. baseline; *p<0.01 vs. CT-1.

Article Snippet: 10− 6 mol/L to inhibit ERK5 (Calbiochem), the STATs inhibitor AG490 at 10−6 mol/L (Calbiochem), antibodies against LIFR (Santa Cruz) and gp130 (Santa Cruz) at 10−6 mol/L.

Techniques: Incubation, Western Blot