메뉴 건너뛰기




Volumn 20, Issue 9, 2010, Pages 2892-2896

Identification of benzimidazole-based inhibitors of the mitogen activated kinase-5 signaling pathway

Author keywords

Benzimidazole; ERK5; Inhibitor; MEK5

Indexed keywords

BENZIMIDAZOLE DERIVATIVE; EPIDERMAL GROWTH FACTOR; MITOGEN ACTIVATED PROTEIN KINASE 7; MITOGEN ACTIVATED PROTEIN KINASE INHIBITOR;

EID: 77950858349     PISSN: 0960894X     EISSN: None     Source Type: Journal    
DOI: 10.1016/j.bmcl.2010.03.033     Document Type: Article
Times cited : (15)

References (41)
  • 30
    • 77950861118 scopus 로고    scopus 로고
    • Jain, P.; Chopra, I.; Yi, S.; Flaherty, P. T.; Madura, J. D. Abstracts of Papers, 238th ACS National Meeting, Washington, DC, United States, August 16-20, 2009, MEDI.
    • Jain, P.; Chopra, I.; Yi, S.; Flaherty, P. T.; Madura, J. D. Abstracts of Papers, 238th ACS National Meeting, Washington, DC, United States, August 16-20, 2009, MEDI.
  • 36
    • 77950861818 scopus 로고    scopus 로고
    • note
    • 3O: C, 73.19; H, 7.17; N, 14.23. Found: C, 73.33; H, 7.03; N, 14.24.
  • 38
    • 77950863165 scopus 로고    scopus 로고
    • HEK293 (ATCC, Manassas, VA) cells were grown on 60 mm culture plates (Sarstedt, Newton, NC) in Dulbecco's Modified Eagle's Medium (DMEM; Gibco, Carlsbad, CA) with 10% heat-inactivated FBS (Atlanta Biological, Lawrenceville, GA, and 0.5% penicillin/streptomycin (Gibco, Carlsbad, CA, Cells were maintained at 37 °C with 5% CO2. The cells were treated with 50 ng/mL epidermal growth factor (EGF; Sigma) 30 min before treatment with the putative MEK5 inhibitors (10 μM of each, Cells were lysed 30 min following treatment with the inhibitors. The cells were washed with 1x PBS and then lysed in 1% Triton X-100 buffer containing 20 mM Tris pH 6.8, 137 mM NaCl, 25 mM beta glycerophosphate, 2 mM NaPPi, 2 mM EDTA, 1 mM Na3 VO4, 10% glycerol, 5 μg/mL leupeptin, 5 μg/mL aprotinin, 2 mM benzamidine, 0.5 mM DTT, and 1 mM PMSF. The lysates were then centrifuged at 10,000 rpm for 10 min at 4 °C. Determination of protein in the supernatant was done usi
    • 4, 10% glycerol, 5 μg/mL leupeptin, 5 μg/mL aprotinin, 2 mM benzamidine, 0.5 mM DTT, and 1 mM PMSF. The lysates were then centrifuged at 10,000 rpm for 10 min at 4 °C. Determination of protein in the supernatant was done using a Bradford assay (Biorad, Hercules, CA).
  • 39
    • 77950864297 scopus 로고    scopus 로고
    • 3 for 1 h and then incubated overnight at 4 °C in primary antibody (1:1000 pERK1/2, 1:500 pERK5, Cell Signaling). Blots were washed with 1x TBS/0.1% Tween for 30 min and then incubated with secondary antibody (1:1000, horseradish peroxidase-conjugated goat anti-rabbit, Upstate). Proteins were visualized with enhanced chemiluminescence (Upstate). Films were scanned and quantified using MATLAB, v.7.1 (Mathworks, Natick, MA).
    • 3 for 1 h and then incubated overnight at 4 °C in primary antibody (1:1000 pERK1/2, 1:500 pERK5, Cell Signaling). Blots were washed with 1x TBS/0.1% Tween for 30 min and then incubated with secondary antibody (1:1000, horseradish peroxidase-conjugated goat anti-rabbit, Upstate). Proteins were visualized with enhanced chemiluminescence (Upstate). Films were scanned and quantified using MATLAB, v.7.1 (Mathworks, Natick, MA).


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.