Journal: Cancer biology & therapy

Article Title: Mismatch repair gene mutations in renal cell carcinoma.

doi: 10.4161/cbt.1.5.171

Figure Lengend Snippet: Figure 1. Western blot analysis to assess hMLH1 and hMSH2 expression in carcinoma cell lines. After fractionation on an 8% SDS-polyacrylamide gel, membranes were incubated with antibodies for MLH1 (top row each panel) or hMSH2 (bottom row each panel). MLH1 polypeptide was not detected in the RCC lines UOB-46 (top panel), STA (middle panel), GET (bottom panel) and the colorectal cell line HCT-116 (top, middle and bottom panels). MSH2 polypeptide is detected in all RCC lines analyzed and absent in the col- orectal cell line LoVo (top, middle and bottom panels).

Article Snippet: Monoclonal antibodies for MLH1(Ab-1), PMS2 (Ab-1) and MSH2 (Ab-1 or 2) were used as directed by the manufacturer (Oncogene Research Products, Cambridge, MA) or as previously described.36 The polyclonal antibody for MSH6 and reagents for detection were used as directed by the manufacturer (Santa Cruz Biotechnology, Santa Cruz, CA).

Techniques: Western Blot, Expressing, Fractionation, Incubation

Journal: Oncogene

Article Title: Interactions of the DNA mismatch repair proteins MLH1 and MSH2 with c-MYC and MAX.

doi: 10.1038/sj.onc.1206252

Figure Lengend Snippet: Figure 2 MLH1 and MSH2 interactions with MYC and MAX detected by GST pull-down assays. (a) In vitro transcribed and translated (IVTT) radiolabelled c-MYC protein. Lane 1 is input c-MYC from IVTT reaction. Fusion proteins used in the pull-down experiments are: lane 2, GST alone; lane 3, Max–GST; lane 4, Mlh1–GST. (b) K562 human promyelocytic cell extracts. Fusion proteins used in the pull-down experiments are: 1, GST alone; 2, Mlh1–F1–GST (Mlh1 amino acids 1–228); 3, Mlh1–F2–GST (Mlh1 amino acids 181–342); 4, Mlh1–F3–GST (Mlh1 amino acids 308–580); 5, Mlh1–F4–GST (Mlh1 amino acids 515–756); 6, Mlh1–GST (full-length Mlh1 amino acids 1–756); 7, Max–GST. Western blots were incubated with c-Myc, Pms2 and Msh2 monoclonal antibodies as shown. (c) A2780 (MLH1 proficient) cell extract. Fusion proteins used in the pull-down experiments are: 1, GST alone; 2, Mlh1–GST; 3, Max–GST. Western blots were incubated with Msh2 monoclonal antibodies. (d) A2780/cp70 (MLH1 deficient) cell extract. Fusion proteins used in the pull-down experiments are: 1, GST alone; 2, Mlh1–GST; 3, Max–GST: Western blots were incubated with Msh2 monoclonal antibodies

Article Snippet: Proteins interacting with the GST fusion proteins were identified using Western blots with antibodies to c-MYC (rabbit polyclonal C19, Santa Cruz, CA, USA), PMS2 (mouse monoclonal Ab1, Calbiochem) and MSH2 (mouse monoclonal Ab2, Calbiochem) or radiolabelled proteins visualized after electrophoresis and autoradiography.

Techniques: In Vitro, Western Blot, Incubation, Bioprocessing