Journal: Environmental toxicology

Article Title: Gallic acid attenuates metastatic potential of human colorectal cancer cells through the miR-1247-3p-modulated integrin/FAK axis.

doi: 10.1002/tox.24087

Figure Lengend Snippet: FIGURE 5 Involvement of miR-1247-3p in the Gallic acid (GA)-inhibited integrin/FAK cascade in DLD-1 cells. Cells were treated with 90 μM GA for 24 h, lysed for total RNA extraction, and subjected to (A) miRNA expression profiling assay via microarray analysis or (B) quantitative analysis of the miRNAs via qPCR. miRNAs with up- and downregulation in response to GA were labeled with red and green, respectively. (C) Cells were transfected with control vector (Ctl-miR) or anti-miR-1247-3p (anti-miR), treated with GA for 24 h, and subjected to the assessment of integrins, FAK, and paxillin by Western blot. Densitometric analysis was performed for semi-quantitation of signals. The tubulin signal was used as internal control. Average signal ratios of phosphorylated protein/total protein compared with the control were indicated.

Article Snippet: Antibodies specifically recognizing human tubulin (SC-134237), integrin αV (SC-376156), integrin β3(SC-365679), focal adhesion kinase (FAK, SC-271126), phosphorylated FAK (pY397-FAK, SC-81493), paxillin (SC-136297), phosphorylated paxillin (pY118-Paxillin, SC-365020), Src (SC-32789), phosphorylated Src (pY416-Src, SC-24621, and CS#2101), and peroxidase-conjugated antibodies against mouse IgG or rabbit IgG were obtained from Santa Cruz Biotechnology, Inc. (CA, USA).

Techniques: RNA Extraction, Expressing, Microarray, Labeling, Transfection, Control, Plasmid Preparation, Western Blot, Quantitation Assay

Journal: Environmental toxicology

Article Title: Gallic acid attenuates metastatic potential of human colorectal cancer cells through the miR-1247-3p-modulated integrin/FAK axis.

doi: 10.1002/tox.24087

Figure Lengend Snippet: FIGURE 4 Gallic acid (GA) reduced integrin expression and inhibited FAK/Paxillin/Src and PI3K/AKT signaling in DLD-1 cells. Cells were treated with GA at the indicated concentrations for 24 h; collected; and lysed for the immunodetection of (A) integrins and the associated signaling proteins, (B) phosphor-paxillin and Src, and (C) PI3K, phosphor-AKT, and AKT via Western blot. Semi-quantitation of signals was conducted by densitometric analysis. DMSO treatment was used as control (C). Densitometric analysis was performed for the semi-quantitation of signals. The tubulin signal was used as internal control. Average signal ratios of phosphorylated protein/total protein compared with the control were indicated.

Article Snippet: Antibodies specifically recognizing human tubulin (SC-134237), integrin αV (SC-376156), integrin β3(SC-365679), focal adhesion kinase (FAK, SC-271126), phosphorylated FAK (pY397-FAK, SC-81493), paxillin (SC-136297), phosphorylated paxillin (pY118-Paxillin, SC-365020), Src (SC-32789), phosphorylated Src (pY416-Src, SC-24621, and CS#2101), and peroxidase-conjugated antibodies against mouse IgG or rabbit IgG were obtained from Santa Cruz Biotechnology, Inc. (CA, USA).

Techniques: Expressing, Immunodetection, Western Blot, Quantitation Assay, Control

Journal: Oncogene

Article Title: Hyperactivation of MEK-ERK1/2 signaling and resistance to apoptosis induced by the oncogenic B-RAF inhibitor, PLX4720, in mutant N-RAS melanoma cells

doi: 10.1038/onc.2010.408

Figure Lengend Snippet: PLX4720-induced MEK-ERK1/2 hyperactivation occurs rapidly and regulates FAK serine 910 phosphorylation. (a) Sbcl2, WM1361A and WM1366 cell lines were treated with DMSO (−), 1 μM PLX4720 or 10 μM U0126 for 16 h. Lysates were analyzed by western blotting (Boisvert-Adamo and Aplin, 2006) using antibodies for (p)FAK-S910 (Biosource Int, Camarillo, CA, USA), (p)FAK-Y397 (Bd Biosciences, San Jose, CA, USA), and total FAK (Cell Signaling Technology). (b) Sbcl2 and WM1366 cells were treated with 1 μM PLX4720 for 15 min to 20 h. Lysates were analyzed by western blot analysis for (p)MEK, MEK, (p)ERK1/2 and ERK1/2.

Article Snippet: Together, these data demonstrate that PLX4720 treatment of mutant N-RAS and wild-type N-RAS/B-RAF melanoma cell lines leads to enhanced activation of the MEK-ERK1/2 pathway. fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window Figure 2 caption a7 PLX4720-induced MEK-ERK1/2 hyperactivation occurs rapidly and regulates FAK serine 910 phosphorylation. (a) Sbcl2, WM1361A and WM1366 cell lines were treated with DMSO (−), 1 μM PLX4720 or 10 μM U0126 for 16 h. Lysates were analyzed by western blotting ( Boisvert-Adamo and Aplin, 2006 ) using antibodies for (p)FAK-S910 (Biosource Int, Camarillo, CA, USA), (p)FAK-Y397 (Bd Biosciences, San Jose, CA, USA), and total FAK (Cell Signaling Technology). (b) Sbcl2 and WM1366 cells were treated with 1 μM PLX4720 for 15 min to 20 h. Lysates were analyzed by western blot analysis for (p)MEK, MEK, (p)ERK1/2 and ERK1/2.

Techniques: Western Blot