HUVEC cells were transiently transfected with 6X DBE-luciferase and pRL-TK plasmids for 24 h 69

After transfection, HUVEC cells were pretreated with AKT inhibitor IV (1 μM) and/or MEK1/2 inhibitor PD98059 (10 μM) for 2 h, followed by treatment with or without EGCG (40 μM) for 24 h. Cells were harvested for firefly/Renilla luciferase assays using the Dual-Luciferase Reporter Assay System (Promega). Luciferase counts were normalized using luciferase transfection control. Data represent the mean ± S.D. *, #, ** = significantly different from respective controls, P < 0.05.<p><b>Copyright information:</b></p><p>Taken from "Inhibition of PI3K/AKT and MEK/ERK pathways act synergistically to enhance antiangiogenic effects of EGCG through activation of FOXO transcription factor"</p><p>http://www.jmolecularsignaling.com/content/3/1/7</p><p>Journal of Molecular Signaling 2008;3():7-7.</p><p>Published online 20 Mar 2008</p><p>PMCID:PMC2278143.</p><p></p

HUVEC cells were transiently transfected with empty vector or constructs encoding FOXO1-TM, FOXO3a-TM, or FOXO4-TM together with 6X DBE-luciferase and pRL-TK plasmids for 24 h 69

Luciferase counts were normalized using luciferase transfection control. After transfection, cells were washed, treated with EGCG (20 μM) for 24 h, and harvested for firefly/Renilla luciferase assays using the Dual-Luciferase Reporter Assay System (Promega). Data represent the mean ± S.D. *, #, ** = significantly different from respective controls, P < 0.05.<p><b>Copyright information:</b></p><p>Taken from "Inhibition of PI3K/AKT and MEK/ERK pathways act synergistically to enhance antiangiogenic effects of EGCG through activation of FOXO transcription factor"</p><p>http://www.jmolecularsignaling.com/content/3/1/7</p><p>Journal of Molecular Signaling 2008;3():7-7.</p><p>Published online 20 Mar 2008</p><p>PMCID:PMC2278143.</p><p></p

Human umbilical vein endothelial cells (HUVECs) were pretreated with AKT inhibitor-IV (1 μM) or MEK1/2 inhibitor PD98059 (10 μM) for 4 h, followed by treatment with EGCG (40 μM) for 2 h

Cells were harvested and TUNEL assay was performed as per manufacturer's instructions (Promega). Data represent mean ± SD. * = significantly different from control, P < 0.05.<p><b>Copyright information:</b></p><p>Taken from "Inhibition of PI3K/AKT and MEK/ERK pathways act synergistically to enhance antiangiogenic effects of EGCG through activation of FOXO transcription factor"</p><p>http://www.jmolecularsignaling.com/content/3/1/7</p><p>Journal of Molecular Signaling 2008;3():7-7.</p><p>Published online 20 Mar 2008</p><p>PMCID:PMC2278143.</p><p></p

Immunohistochemical examination of Bcl-2 family members and death receptors

<p><b>Copyright information:</b></p><p>Taken from "Curcumin sensitizes TRAIL-resistant xenografts: molecular mechanisms of apoptosis, metastasis and angiogenesis"</p><p>http://www.molecular-cancer.com/content/7/1/16</p><p>Molecular Cancer 2008;7():16-16.</p><p>Published online 29 Jan 2008</p><p>PMCID:PMC2249593.</p><p></p> Immunohistochemistry was performed to measure the expression of Bak, Bax, Bcl-2, Bcl-X, TRAIL-R1/DR4 and TRAIL-R2/DR5 in tumor tissues derived from control and/or treated mice on week 6

Effects of STAT3 shRNA on the regulation of cyclin D1, Bcl-X_L and c-Myc by EGCG.

<p>(A), AsPC-1/scrambled and AsPC-1/STAT3 shRNA cells were treated with or without EGCG (60 µM) for 48 h. The expression of cyclin D1 was measured by q-RT-PCR. Data represent mean ± SD. * = significantly different from respective controls, P<0.05. (B), PANC-1/scrambled and PANC-1/STAT3 shRNA cells were treated with or without EGCG (60 µM) for 48 h. The expression of cyclin D1 was measured by q-RT-PCR. Data represent mean ± SD. * = significantly different from respective controls, P<0.05. (C), PANC-1/scrambled and PANC 1/STAT3 shRNA cells were treated with or without EGCG (60 µM) for 48 h. The expression of Bcl-X_L was measured by qRT-PCR. Data represent mean ± SD. * = significantly different from respective controls, P<0.05. (D), PANC-1/scrambled and PANC-1/STAT3 shRNA cells were treated with or without EGCG (60 µM) for 48 h. The expression of c-Myc was measured by q-RT-PCR. Data represent mean ± SD. * = significantly different from respective controls, P<0.05.</p

Effects of curcumin and/TRAIL on markers of phospho-p65NFκB, Cox-2, and IL-8

<p><b>Copyright information:</b></p><p>Taken from "Curcumin sensitizes TRAIL-resistant xenografts: molecular mechanisms of apoptosis, metastasis and angiogenesis"</p><p>http://www.molecular-cancer.com/content/7/1/16</p><p>Molecular Cancer 2008;7():16-16.</p><p>Published online 29 Jan 2008</p><p>PMCID:PMC2249593.</p><p></p> (A), Immunohistochemistry was performed to measure the expression of phospho-p65NFκB, Cox-2 and IL-8 in tumor tissues derived from control and/or treated mice on week 6. (B), Expression of phospho-p65NFκB, Cox-2, IL-8 and β-actin in tumor tissues derived on week 6 were measured by the Western blot analysis. (C), NFκB-DNA binding activity. Nuclear extracts were prepared from tumor tissues derived from different treatment groups on week 6. NFκB-DNA binding activity was measured by Gelshift assay as described in Materials and Methods. The relative nuclear NFκB-DNA binding activities were quantified by scanning densitometry

Schematic representation of the inhibition of SHH signaling and genes involved in the balance between cellular proliferation and cell death.

<p>Activated Gli1 and Gli2, downstream of SHH-Patched-Smoothened, regulate targets of SHH signaling including Bcl-2, PDGFRα, Fas, and DRs. GDC-0449 (targeting Smoothened) blocks the indirect functions of Gli activators, resulting in cell death.</p

Impact of SHH signaling pathway on the regulation of cell survival and antiproliferative effects of GDC-0449.

<p>(A), Knockout of Gli1 shRNA and Gli2 shRNA in human pancreatic CSCs. Pancreatic CSCs were transduced with lentiviral particles expressing Scrambled, Gli1 shRNA, Gli2 shRNA or Gli1 plus Gli2 shRNA (KO). (B), Pancreatic CSCs were treated with GDC-0449 (0, 1, 5 and 10 µM) for 72 h, and cell viability and apoptosis was measured in scrambled and Gli1 plus Gli2 shRNA CSCs. Data represent mean ± SD, n = 4. @ or # significantly different from respective control (P<0.05). (C), Scrambled and Gli1 plus Gli2 shRNA pancreatic CSCs were treated with GDC-0449 (0 and 10 µM) for 48 h, and lysates were extracted to determine the expression of DR4, DR5, PDGFRα, Fas and Bcl-2 by Western blot analysis. β-Actin was used as the loading control. (D), Inhibition of primary and secondary spheroids by GDC-0449. Pancreatic CSCs (scrambled, and Gli1 + Gli2 shRNA) were seeded in suspension and treated with GDC-0449 (10 µM) for 7 days. At the end of incubation period, spheroids were collected, and dissociated with Accutase (Innovative Cell Technologies, Inc.). For secondary spheroids, cells were reseeded and treated with GDC-0449 (10 µM) for additional 7 days. Cell viability was measured by trypan blue assay. Data represent mean ± SD. @ or # significantly different from respective controls, P<0.05.</p

Inhibition of STAT3 enhances the inhibitory effects of EGCG on motility and cell viability of pancreatic cancer cells.

<p>(A), AsPC-1 and PANC-1 were transfected with STAT3 shRNA. The expression of STAT3 was performed by Western blotting. (B), AsPC-1 scratch assay. AsPC-1 scrambled and STAT3 shRNA cells were cultured in 6 well dishes. The scratch was marked when the dishes were 50% confluent. Pictures were taken after the cells were treated with EGCG and incubated for 0, 24 and 48 h. (C), Cell viability assay. AsPC-1 and PANC-1 (scrambled and STAT3 shRNA) cells were seeded and treated with EGCG (0, 20, 40, 60 µM). After 72 h of treatment, cell viability was performed by XTT assay. Data represent mean ± SD. * or ** = significantly different from respective controls, P<0.05.</p

EGCG and gemcitabine inhibit cell viability and STAT3 target genes.

<p>(A), AsPC-1 and PANC-1 cells were treated with EGCG (0, 20, 40, 60 µM) with or without gemcitabine (0.5 µM) for 72 h. Cell viability was measured by XTT assay. Data represent mean ± SD. * or ** = significantly different from respective controls, P<0.05. (B), AsPC-1 and PANC-1 cells were treated with EGCG (0, 20, 40, 60 µM) with or without gemcitabine (0.5 µM) for 72 h. Apoptosis was measured by TUNEL assay. Data represent mean ± SD. * or ** = significantly different from respective controls, P<0.05. (C), Inhibition of STAT3 target genes by EGCG and gemcitabine. AsPC-1 and PANC-1 cells were treated with EGCG (20 µM) or gemcitabine (0.5 µM) for 48 h. The expression of VEGF, c-Myc, survivin and cyclin D1was was measured by qRT-PCR. Data represent mean ± SD. * = significantly different from respective controls, P<0.05.</p