Journal: International journal of oncology

Article Title: JAK/STAT3 signaling is required for TGF-β-induced epithelial-mesenchymal transition in lung cancer cells.

doi: 10.3892/ijo.2014.2310

Figure Lengend Snippet: Figure 2. AG490 inhibits Smad3 activation and transcriptional responses in lung cancer A549 and H1650 cells. (A) AG490 inhibited phosphorylation of Stat3. After serum starvation overnight, cells were subjected to AG490 treatment (JAK2/STAT3 inhibitor, 50 or 100 µM) for 4 h, and the expression of p-Stat3, total Stat3 was analyzed using western blotting. (B) AG490 suppressed TGF-β-induced Smad3 activation and Snail upregulation. Cells were pretreated with 50 µM AG490 for 4 h and then treated with 5 ng/ml TGF-β1 for 4 h as indicated. Levels of p-Samd3, Smad3 and Snail were monitored by western blotting. (C) AG490 attenuated TGF-β-induced PAI-1 promoter activation. After transient transfection with PAI-1 promoter construct, cells were treated with 50 µM AG490 for 4 h and then incubated for 18 h in the absence or presence of 5 ng/ml TGF-β1. Relative luciferase activity was expressed as the mean fold change from basal level ± SD of three independent experiments. (D and E) AG490 diminished TGF-β-induced increase in Snail and MMP2. Cells were treated with 50 µM AG490 for 4 h and then exposed to 5 ng/ml TGF-β1, and the mRNA levels of Snail (treated with TGF-β1 for 1 h) and MMP2 (24 h) were determined by quantitative RT-PCR. *P<0.05; **P<0.01.

Article Snippet: JAK2 inhibitor AG490 and Smad3 phosphorylation inhibitor SIS3 were purchased from Merck KGaA (Darmstadt, Germany) and Santa Cruz Biotechnology (Santa Cruz, CA, USA), respectively.

Techniques: Activation Assay, Phospho-proteomics, Expressing, Western Blot, Transfection, Construct, Incubation, Luciferase, Activity Assay, Quantitative RT-PCR

Journal: International journal of oncology

Article Title: JAK/STAT3 signaling is required for TGF-β-induced epithelial-mesenchymal transition in lung cancer cells.

doi: 10.3892/ijo.2014.2310

Figure Lengend Snippet: Figure 3. AG490 abrogates TGF-β-induced migration and invasion in A549 and H1650 cells. (A) AG490 inhibited TGF-β-induced cell migration. Cells were treated with 50 µM AG490 for 4 h followed by 5 ng/ml TGF-β1 for 12 h, and allowed to migrate through an 8-µM pore in transwells. Migrated cells through the pores were stained with 1% crystal violet and counted under a light microscope (magnification, x200). (B) AG490 suppressed TGF‑β-induced cell invasion. Cells were treated as above and allowed to pass through Matrigel-coated membrane in transwells. Invaded cells through the filter were stained and counted. The data summarized in the bar charts are presented as mean ± SD of three independent fields. *P<0.05; **P<0.01.

Article Snippet: JAK2 inhibitor AG490 and Smad3 phosphorylation inhibitor SIS3 were purchased from Merck KGaA (Darmstadt, Germany) and Santa Cruz Biotechnology (Santa Cruz, CA, USA), respectively.

Techniques: Migration, Staining, Light Microscopy, Membrane

Journal: BMC Complementary Medicine and Therapies

Article Title: Potentilla reptans L. postconditioning protects reperfusion injury via the RISK/SAFE pathways in an isolated rat heart

doi: 10.1186/s12906-021-03456-2

Figure Lengend Snippet: Experimental protocols. All experimental groups were first perfused for 30 mins on the Langendorff apparatus to allow the isolated hearts to stabilize. The hearts were then divided into different groups. All groups were subjected to 30 mins of regional ischemia followed by 100 mins of reperfusion. IR; Ischemia-reperfusion, IPOST; Ischemic postconditioning (3 × 10 s ischemia and reperfusion at the onset of reperfusion period), Etpost; ethyl acetate fraction from Potentilla reptans root (2 μg/ml) was applied at the onset of reperfusion, L-NAME; NO inhibitor (50 μM), Wort; Wortmannin a PI3K/AKT inhibitor (400 nM), PD; PD98059 (2′-Amino-3′-methoxyflavone) an ERK1/2 inhibitor (400 nM), AG; tyrphostin (AG490, 400 nM) a JAK/STAT3 inhibitor, HD; 5-hydroxy decanoate (5HD,1 μM) a mitoKATP channel blocker

Article Snippet: Wortmannin (Wort; the PI3k/Akt inhibitor), PD98059 (PD; the ERK1/2 inhibitor), Tyrphostin AG490 (the JAK/STAT3 inhibitor), and 5-hydroxy decanoate (a mitoKATP channel blocker) were purchased from “Santa Cruz Biotech, Dallas, Texas, USA”.

Techniques: Isolation

Journal: BMC Complementary Medicine and Therapies

Article Title: Potentilla reptans L. postconditioning protects reperfusion injury via the RISK/SAFE pathways in an isolated rat heart

doi: 10.1186/s12906-021-03456-2

Figure Lengend Snippet: Western blot analysis. A Western blot analysis shows the relative intensity of AKT (Tyr473) phosphorylation in heart tissue of IR, Etpost (2 μg/ml) and Etpost+Wort groups that were adjusted relative to GAPDH. B Western blot analysis illustrates the relative intensity of ERK1/2 (Thr183/185) phosphorylation in heart tissue of IR, Etpost (2 μg/ml) and Etpost+PD groups that were adjusted relative to GAPDH. C Western blot analysis displays the relative intensity of STAT3 (Tyr705) phosphorylation in heart tissue of IR, Etpost (2 μg/ml) and Etpost+AG groups that were adjusted relative to GAPDH. The data were expressed as mean ± SEM. * P < 0.05 vs. IR group and # P < 0.05 vs. Etpost (2 μg/ml). Etpost: ethyl acetate fraction of P. reptans root; Wort: Wortmanin (PI3K/Akt inhibitor); PD: PD98059 (ERK inhibitor); AG: AG490 (JAK/STAT3 inhibitor)

Article Snippet: Wortmannin (Wort; the PI3k/Akt inhibitor), PD98059 (PD; the ERK1/2 inhibitor), Tyrphostin AG490 (the JAK/STAT3 inhibitor), and 5-hydroxy decanoate (a mitoKATP channel blocker) were purchased from “Santa Cruz Biotech, Dallas, Texas, USA”.

Techniques: Western Blot, Phospho-proteomics

Journal: Biochimica et biophysica acta

Article Title: Prolactin and glucocorticoid signaling induces lactation-specific tight junctions concurrent with β-casein expression in mammary epithelial cells.

doi: 10.1016/j.bbamcr.2016.04.023

Figure Lengend Snippet: Fig. 4. Prl and Dex influence the expression patterns of Cldn3 and Cldn4 in MECs (A) Immunostaining images of Cldn3 (green) and Cldn4 (green) with Ocln (red) in MECs cultured with or without Prl (0.5 U/mL) and Dex (1 μM) for 5 days. Nuclei stained with DAPI are blue. The scale bars represent 10 μm. (B–F) The results of western blotting and densitometry analysis of Cldn3 and Cldn4 with β-actin used as an internal control. MECs were cultured for 5 days with or without Prl (0.5 U/mL or 0–2.5 U/mL) and Dex (1 μM). MECs were pretreated with 10 μM AG490 (JAK inhibitor) and 5 μM RU486 (glucocorticoid receptor antagonist) 1 h before Prl (0.5 U/mL) and Dex (100 nM) treatment. The data are presented as the mean (SEM); n = 5; *p b 0.05 and **p b 0.005. (G) The expression of Cldn3 and Cldn4, determined by qRT-PCR. The data are presented as the mean (SEM); n = 6; *p b 0.05 and **p b 0.005.

Article Snippet: AG490 (JAK inhibitor; cat. no. T9969) and RU486 (GR antagonist; cat. no. M3321)were purchased from LKT Laboratories, Inc. (St. Paul, MN).

Techniques: Expressing, Immunostaining, Cell Culture, Staining, Western Blot, Control, Quantitative RT-PCR