Journal: Oncology Reports

Article Title: CAFs enhance paclitaxel resistance by inducing EMT through the IL-6/JAK2/STAT3 pathway

doi: 10.3892/or.2018.6311

Figure Lengend Snippet: The JAK2/STAT3 pathway is required for regulating EMT in ovarian cancer cells induced by IL-6. (A) The culture supernatants of CAFs and NFs were applied to OVCAR3 cells. The phosphorylation levels of JAK2 and STAT3 in OVCAR3 cells treated with CAF supernatant were significantly higher than those in cells treated with NF supernatant. After the addition of IL-6 mAb, the phosphorylation levels of JAK2 and STAT3 were decreased. (B) After the JAK2/STAT3-signaling-pathway-specific inhibitor AG490 was added, the expression of the interstitial markers N-cadherin and Vimentin was decreased and the expression of the epithelium marker E-cadherin was increased. These results indicated that CAF-derived IL-6 could mediate EMT in OVCAR3 cells via the JAK2/STAT3 pathway.

Article Snippet: The JAK2/STAT3 pathway inhibitor AG490 was purchased from APExBIO (Apexbio Technology LLC, Houston, TX, USA) and the β-TGF inhibitor SB431542 was obtained from Selleck Chemicals (Houston, TX, USA).

Techniques: Phospho-proteomics, Expressing, Marker, Derivative Assay

Journal: Oncology Reports

Article Title: CAFs enhance paclitaxel resistance by inducing EMT through the IL-6/JAK2/STAT3 pathway

doi: 10.3892/or.2018.6311

Figure Lengend Snippet: CAF-derived IL-6 enhances paclitaxel resistance of ovarian cancer cells through cellular EMT. (A) The culture supernatants of CAFs and NFs were applied to OVCAR3 cells. The number of apoptotic cells was decreased in OVCAR3 cells treated with CAF supernatant. After the addition of IL-6 mAb, paclitaxel resistance was reduced and paclitaxel-induced apoptosis was promoted. (B) The expression of pro-apoptotic protein Bax and caspase-3-p17 was decreased, and the expression of apoptosis-suppressing protein Bcl-2 was enhanced in cells treated with CAF supernatant compared with NF supernatant. (C) The number of apoptotic cells treated with paclitaxel was increased after the addition of SB431542 and AG490.

Article Snippet: The JAK2/STAT3 pathway inhibitor AG490 was purchased from APExBIO (Apexbio Technology LLC, Houston, TX, USA) and the β-TGF inhibitor SB431542 was obtained from Selleck Chemicals (Houston, TX, USA).

Techniques: Derivative Assay, Expressing

Journal: Acta pharmacologica Sinica

Article Title: Berberine protects rat heart from ischemia/reperfusion injury via activating JAK2/STAT3 signaling and attenuating endoplasmic reticulum stress.

doi: 10.1038/aps.2015.136

Figure Lengend Snippet: Figure 2. BBR or AG490 treatment had no significant toxic effect on sham-operated rat hearts. Normal rats pretreated with BBR or AG490 were subjected to the sham operation. After 24 h of reperfusion, echocardiography was performed. After 6 h of reperfusion, the myocardial infarct size and apoptosis index were evaluated. (A) Representative M-mode echocardiographic images. (B) Left ventricular ejection fraction (LVEF). (C) Left ventricular fractional shortening (LVFS). (D) Representative heart section images. The Evans blue-stained areas (blue) indicate the non-ischemic/reperfused area; the TTC-stained areas (red) indicate ischemic but viable tissue; and the Evans blue/TTC-unstained (negative) areas (white) indicate infarcted myocardium. (E) Representative images of apoptotic cardiomyocytes by TUNEL staining. The green fluorescence shows the TUNEL-positive nuclei; the blue fluorescence shows the nuclei of all cardiomyocytes; original magnification, ×400. (F) The myocardial infarct size expressed as the percentage of area-at-risk. (G) Percentage of TUNEL-positive nuclei. (H) Representative blots. (I) p-JAK2/JAK2 ratio. (J) p-STAT3/STAT3 ratio. BBR, berberine; AG, AG490. The results are expressed as the mean±SEM. n=8/group. cP<0.01 vs the sham group; fP<0.01 vs the sham+BBR group.

Article Snippet: The primary antibodies against JAK2, p-JAK2 (Tyr1007/1008), STAT3, p-STAT3 (Try705), p-PERK (Thr981), PERK, p-eIF2α (Ser52), eIF2α, ATF4, CHOP, gp91phox, caspase-3, Bcl-2, Bax, and β-actin, AG490 (the specific inhibitor of JAK2/STAT3 signaling), and JAK2 siRNA were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Staining, TUNEL Assay, Fluorescence

Journal: Acta pharmacologica Sinica

Article Title: Berberine protects rat heart from ischemia/reperfusion injury via activating JAK2/STAT3 signaling and attenuating endoplasmic reticulum stress.

doi: 10.1038/aps.2015.136

Figure Lengend Snippet: Figure 3. BBR effectively reduced the MI/R injury, whereas AG490 treatment blocked this effect. Normal rats pretreated with BBR or AG490 were subjected to the MI/R operation. After 24 h of reperfusion, echocardiography was performed. After 6 h of reperfusion, the myocardial infarct size and apoptotic index were evaluated. (A) Representative M-mode echocardiographic images. (B) Left ventricular ejection fraction (LVEF). (C) Left ventricular fractional shortening (LVFS). (D) Representative images of apoptotic cardiomyocytes by TUNEL staining. The green fluorescence shows the TUNEL- positive nuclei; the blue fluorescence shows the nuclei of all cardiomyocytes; original magnification, ×400. (E) Percentage of TUNEL-positive nuclei. (F) Representative heart section images. The Evans blue-stained areas (blue) indicate the non-ischemic/reperfused area; the TTC-stained areas (red) indicate ischemic but viable tissue; and the Evans blue/TTC-unstained (negative) areas (white) indicate infarcted myocardium. (G) The myocardial infarct size expressed as the percentage of area-at-risk. BBR, berberine; AG, AG490. The results are expressed as the mean±SEM. n=8/group.

Article Snippet: The primary antibodies against JAK2, p-JAK2 (Tyr1007/1008), STAT3, p-STAT3 (Try705), p-PERK (Thr981), PERK, p-eIF2α (Ser52), eIF2α, ATF4, CHOP, gp91phox, caspase-3, Bcl-2, Bax, and β-actin, AG490 (the specific inhibitor of JAK2/STAT3 signaling), and JAK2 siRNA were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: TUNEL Assay, Staining, Fluorescence

Journal: Acta pharmacologica Sinica

Article Title: Berberine protects rat heart from ischemia/reperfusion injury via activating JAK2/STAT3 signaling and attenuating endoplasmic reticulum stress.

doi: 10.1038/aps.2015.136

Figure Lengend Snippet: Figure 4. BBR effectively ameliorated cardiac necrosis and oxidative stress, whereas AG490 treatment blocked these effects. Normal rats treated with BBR or AG490 were subjected to the MI/R operation. All measurements were performed after 6 h of reperfusion. (A) Serum LDH levels. (B) Serum CK levels. (C) Cardiac superoxide generation. (D) gp91phox expression. Top images: representative blots. (E) Myocardial MDA contents. (F) Myocardial SOD contents. BBR, berberine; AG, AG490. The results are expressed as the mean±SEM. n=8/group. cP<0.01 vs the MI/R+V group; fP<0.01 vs the MI/R +BBR group; iP<0.01 vs the MI/R+BBR+AG group.

Article Snippet: The primary antibodies against JAK2, p-JAK2 (Tyr1007/1008), STAT3, p-STAT3 (Try705), p-PERK (Thr981), PERK, p-eIF2α (Ser52), eIF2α, ATF4, CHOP, gp91phox, caspase-3, Bcl-2, Bax, and β-actin, AG490 (the specific inhibitor of JAK2/STAT3 signaling), and JAK2 siRNA were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Expressing

Journal: Acta pharmacologica Sinica

Article Title: Berberine protects rat heart from ischemia/reperfusion injury via activating JAK2/STAT3 signaling and attenuating endoplasmic reticulum stress.

doi: 10.1038/aps.2015.136

Figure Lengend Snippet: Figure 5. BBR up-regulated cardiac JAK2/STAT3 signaling and down-regulated myocardial apoptosis, whereas AG490 treatment blocked these effects. Normal rats pretreated with BBR or AG490 were subjected to the MI/R operation. All measurements were performed after 4 h of reperfusion. (A) Representative blots. (B) p-JAK2/JAK2 ratio. (C) p-STAT3/STAT3 ratio. (D) Caspase-3 expression. (E) Bcl-2 expression. (F) Bax expression. BBR, berberine; AG, AG490. The results are expressed as the mean±SEM. n=8/group. cP<0.01 vs the MI/R+V group; fP<0.01 vs the MI/R+BBR group;

Article Snippet: The primary antibodies against JAK2, p-JAK2 (Tyr1007/1008), STAT3, p-STAT3 (Try705), p-PERK (Thr981), PERK, p-eIF2α (Ser52), eIF2α, ATF4, CHOP, gp91phox, caspase-3, Bcl-2, Bax, and β-actin, AG490 (the specific inhibitor of JAK2/STAT3 signaling), and JAK2 siRNA were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Expressing

Journal: Acta pharmacologica Sinica

Article Title: Berberine protects rat heart from ischemia/reperfusion injury via activating JAK2/STAT3 signaling and attenuating endoplasmic reticulum stress.

doi: 10.1038/aps.2015.136

Figure Lengend Snippet: Figure 6. BBR significantly attenuated the PERK/eIF2α/ATF4-mediated myocardial ER stress, whereas AG490 treatment blocked this effect. Normal rats pretreated with BBR or AG490 were subjected to the MI/R operation. Western blotting analysis was performed after 4 h of reperfusion. (A) Representative blots; (B) p-PERK/PERK ratio; (C) p-eIF2α/eIF2α ratio; (D) ATF4 expression; (E) CHOP expression. BBR, berberine; AG, AG490. The results are expressed as the mean±SEM. n=8/group. cP<0.01 vs the MI/R+V group; eP<0.05, fP<0.01 vs the MI/R+BBR group; iP<0.01 vs the MI/R +BBR+AG group.

Article Snippet: The primary antibodies against JAK2, p-JAK2 (Tyr1007/1008), STAT3, p-STAT3 (Try705), p-PERK (Thr981), PERK, p-eIF2α (Ser52), eIF2α, ATF4, CHOP, gp91phox, caspase-3, Bcl-2, Bax, and β-actin, AG490 (the specific inhibitor of JAK2/STAT3 signaling), and JAK2 siRNA were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Western Blot, Expressing