Journal: International Journal of Molecular Sciences

Article Title: Evodiamine Induces Apoptosis and Inhibits Migration of HCT-116 Human Colorectal Cancer Cells

doi: 10.3390/ijms161126031

Figure Lengend Snippet: Effects of AG490 and EVO on PGI, the JAK2/STAT3 signaling pathway, and MMP3 in HCT-116 cells. ( A , C ) HCT-116 cells were treated with various concentrations of AG490 for 48 h and then stimulated with 25 ng/mL IL-6 for 15 min, after which whole-cell extracts of PGI, MMP3, JAK2, p-JAK2 (data not shown), STAT3, and p-STAT3 were analyzed by Western blotting; ( E ) HCT-116 cells were treated with 50 μmol/L AG490, 6 μmol/L EVO, and combined 50 μmol/L AG490 with 6 μmol/L EVO for 6 h, then stimulated with 25 ng/mL IL-6 for 15 min. Protein expression was analyzed by Western blotting; ( B , D , F ) The graph represents densitometry of the results of three independent experiments (mean ± SD). * p < 0.05, ** p < 0.01 compared to the control. The control group of A–C with 25 ng/mL IL-6 + 0 μmol/L AG490, while E with 25 ng/mL IL-6 + 0 μmol/L AG490 + 0 μmol/L EVO.

Article Snippet: AG490 was obtained from Tocris Bioscience (Bristol, UK).

Techniques: Western Blot, Expressing, Control

Journal: Scientific reports

Article Title: Interleukin 22 attenuated angiotensin II induced acute lung injury through inhibiting the apoptosis of pulmonary microvascular endothelial cells.

doi: 10.1038/s41598-017-02056-w

Figure Lengend Snippet: Figure 2. IL-22 attenuated the ALI induced by AngII. (A) The incidence of lung injury showed remarkable decrease after IL-22 treatment. (B) In the ALI model induced by AngII, obvious oedema was noticed, together with massive infiltration of neutrophils (MPO) and macrophages (CD68), which was significantly reversed after IL-22 treatment. (C) IL22 could significantly attenuate the lung injury as revealed by pathological changes, but such phenomenon was completely reserved after AG490. HE staining and immunohistochemistry images were observed under a magnification of 200× and 400×, respectively. *P < 0.05 versus control group. #P < 0.05 versus AngII group and AngII+IL-22+AG490 group.

Article Snippet: Besides, AG490 (10 mg/ Kg, sc-202046, Santa Cruz) was given to inhibit the activity of JAK2 every two days via peritoneal injection.

Techniques: Staining, Immunohistochemistry, Control

Journal: Scientific reports

Article Title: Interleukin 22 attenuated angiotensin II induced acute lung injury through inhibiting the apoptosis of pulmonary microvascular endothelial cells.

doi: 10.1038/s41598-017-02056-w

Figure Lengend Snippet: Figure 3. IL22 inhibited the apoptosis of PMVECs induced by AngII. (A) IL22 could attenuate the apoptosis of PMVECs significantly as revealed by TUNEL assay, while such phenomenon was completely inhibited after AG490. (B) Flow cytometry indicated the apoptosis rate of the PMVECs in the AngII+IL22 group was significantly decreased compared with the AngII group. The apoptosis rate in the AngII+IL22+AG490 group was significantly elevated compared with that of AngII+IL22 group. The raw data were listed in the Supplementary file. (C) The expression of Bcl-2 in the PMVECs significantly decreased after AngII stimulation, while the expression of Bcl-2 in the AngII+IL22 group was higher than that of AngII group. *P < 0.05 versus control group. #P < 0.05 versus AngII group and AngII+IL-22+AG490 group. Immunofluorescence images were observed under a magnification of 200×.

Article Snippet: Besides, AG490 (10 mg/ Kg, sc-202046, Santa Cruz) was given to inhibit the activity of JAK2 every two days via peritoneal injection.

Techniques: TUNEL Assay, Flow Cytometry, Expressing, Control, Immunofluorescence

Journal: Scientific reports

Article Title: Interleukin 22 attenuated angiotensin II induced acute lung injury through inhibiting the apoptosis of pulmonary microvascular endothelial cells.

doi: 10.1038/s41598-017-02056-w

Figure Lengend Snippet: Figure 5. IL22 contributed to the expression of STAT3 in the PMVECs. (A) IL22 induced up-regulation of STAT3 in a time-dependent manner, and reached the peak level at 48 h. (B) The expression of STAT3 in the AngII+IL22 group was higher than the other groups. The raw data were listed in the Supplementary file. *P < 0.05 versus control group; #P < 0.05 versus AngII group and AngII+IL-22+AG490 group.

Article Snippet: Besides, AG490 (10 mg/ Kg, sc-202046, Santa Cruz) was given to inhibit the activity of JAK2 every two days via peritoneal injection.

Techniques: Expressing, Control

Journal: Scientific reports

Article Title: Interleukin 22 attenuated angiotensin II induced acute lung injury through inhibiting the apoptosis of pulmonary microvascular endothelial cells.

doi: 10.1038/s41598-017-02056-w

Figure Lengend Snippet: Figure 4. IL22 contributed to the expression of STAT3 in the lung tissues in mice. Immunohistochemistry (A, B) indicated IL22 induced up-regulation of STAT3 in the lung tissues, but such fact was inhibited after AG490. The raw data were listed in the Supplementary file. (C) Western blot analysis indicated the expression of STAT3 in the lung tissues in the AngII+IL22 group was higher than the other groups. *P < 0.05 versus control group; #P < 0.05 versus AngII group and AngII+IL-22+AG490 group. The images of immunohistochemistry were observed under a magnification of 400× and 1000×, respectively.

Article Snippet: Besides, AG490 (10 mg/ Kg, sc-202046, Santa Cruz) was given to inhibit the activity of JAK2 every two days via peritoneal injection.

Techniques: Expressing, Immunohistochemistry, Western Blot, Control

Journal: Scientific reports

Article Title: Interleukin 22 attenuated angiotensin II induced acute lung injury through inhibiting the apoptosis of pulmonary microvascular endothelial cells.

doi: 10.1038/s41598-017-02056-w

Figure Lengend Snippet: Figure 6. IL-22 significantly stimulated phosphorylation at tyrosine 705 of STAT3 in PMVECs. (A) The expression of pY705-STAT3 was up-regulated after IL22 interference, and reached the peak level at 30 min. (B) Immunofluorescence analysis indicated the expression of pY705-STAT3 significantly increased together with accumulation in the nucleus after IL22 interference. (C) The expression of pY705-STAT3 in the AngII+IL22 group was higher than the other groups. The raw data of Western blot were listed in the Supplementary file. *P < 0.05 versus control group; #P < 0.05 versus AngII group and AngII+IL-22+AG490 group.

Article Snippet: Besides, AG490 (10 mg/ Kg, sc-202046, Santa Cruz) was given to inhibit the activity of JAK2 every two days via peritoneal injection.

Techniques: Phospho-proteomics, Expressing, Immunofluorescence, Western Blot, Control

Journal: Scientific reports

Article Title: Interleukin 22 attenuated angiotensin II induced acute lung injury through inhibiting the apoptosis of pulmonary microvascular endothelial cells.

doi: 10.1038/s41598-017-02056-w

Figure Lengend Snippet: Figure 7. IL-22 significantly stimulated nuclear STAT3 expression in PMVECs. (A) STAT3 was mainly localized in the cytoplasm in the control group and AngII group. The expression of STAT3 in the nucleus in the AngII+IL22 group significantly increased, while its expression in the nucleus significantly decreased in the AngII+IL22+AG490 group. (B) Western blot analysis indicated the STAT3 in the nucleus in the AngII+IL22 group was significantly higher than the other groups. The raw data were listed in the Supplementary file. *P < 0.05 versus control group; #P < 0.05 versus AngII+IL-22+AG490 group.

Article Snippet: Besides, AG490 (10 mg/ Kg, sc-202046, Santa Cruz) was given to inhibit the activity of JAK2 every two days via peritoneal injection.

Techniques: Expressing, Control, Western Blot

Journal: iScience

Article Title: Inhibition of CD44 suppresses the formation of fibrotic scar after spinal cord injury via the JAK2/STAT3 signaling pathway

doi: 10.1016/j.isci.2024.108935

Figure Lengend Snippet: STAT3 was the downstream of JAK2 in CD44-mediated fibrotic effect in the inflammatory microenvironment (A) Western blotting analyzed the expression of p -JAK2, p-STAT3, STAT3, and p -Smad2 in the fibroblasts stimulated with IS after inhibition of AG490. (B‒E) Quantification of western blotting results in A. N = 3 of each group by one-way ANOVA. (F) Western blotting analyzed the expression of CD44, α-SMA, collagen-I, FN, and laminin in the fibroblasts stimulated with IS after inhibition of AG490. (G‒K) Quantification of western blotting results in F. N = 3 of each group by one-way ANOVA. (L and M) Quantification analysis of the rate of proliferation (%) (L) per 10 6 μm 2 (n = 5) and cell numbers of migration (M) per 10 6 μm 2 (n = 5) in different groups by one-way ANOVA. All data were presented as mean ± SEM. (∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001). (See also Figure S3 ).

Article Snippet: AG490 , Targetmol , Cat#T2600;CAS:133550-30-8;Batch:147766.

Techniques: Western Blot, Expressing, Inhibition, Migration

Journal: iScience

Article Title: Inhibition of CD44 suppresses the formation of fibrotic scar after spinal cord injury via the JAK2/STAT3 signaling pathway

doi: 10.1016/j.isci.2024.108935

Figure Lengend Snippet:

Article Snippet: AG490 , Targetmol , Cat#T2600;CAS:133550-30-8;Batch:147766.

Techniques: Recombinant, In Vivo, Lysis, Acid Assay, Software

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