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

Journal: Cell Death & Disease

Article Title: Serglycin induces osteoclastogenesis and promotes tumor growth in giant cell tumor of bone

doi: 10.1038/s41419-021-04161-1

Figure Lengend Snippet: A − D Western blot analysis of phosphorylated FAK protein level in RAW264.7 cells after treatment with CM from GCTB-1 ( A ) or GCTB-19 ( B ) with SRGN knockdown, CM from hFOB1.19 with SRGN overexpression ( C ), or human recombinant SRGN protein ( D ). E Western blotting analysis of phosphorylated FAK protein level in RAW264.7 cells after treatment with CM from hFOB1.19 with SRGN overexpression and the CD44 neutralizing antibody. F Western blotting analysis of phosphorylated FAK in RAW264.7 cells after treatment with human recombinant SRGN protein and the CD44 neutralizing antibody. G , H Western blotting analysis of phosphorylated FAK in RAW264.7 CD44 -knockout cells after treatment with CM from hFOB1.19 with SRGN overexpression ( G ) and human recombinant SRGN protein ( H ). I FAK phosphorylation levels in human GCTB samples with different levels of SRGN expression. Protein expression was scored as 0 (negative), 1 (weak), 2 (moderate) and 3 (strong) by immunohistochemistry staining ( n = 71 patients). Scale bar, 100 μm. P values were obtained by chi-squared test ( I ).

Article Snippet: The antibodies used for Western blotting, immunoprecipitation and immunohistochemistry were as follows: β-ACTIN (A2228, Sigma), Flag (F1804, Sigma), SRGN (sc-374657, Santa Cruz), CD44 (37259, CST), His (12698, CST), FAK (A11531, Abclonal), phosphor-FAK (AP0302, Abclonal), H3.3 G34W (RM263, RevMAb).

Techniques: Western Blot, Over Expression, Recombinant, Knock-Out, Expressing, Immunohistochemistry, Staining