Transcription profiling of mouse 6780 cell line derived from murine T-cell lymphomaTGFbeta dependent gene expression in T-cell lymphomas

ABSTRACT: The cell line "6780" derived from murine T-cell lymphoma was grown in RPMI medium 1640 with 10% FBS/1% penicillin/Streptomycin/1% l-glutamine/50 μM 2-mercaptoethanol. Vector constructs containing a dominant negative mutant of the human TGF-beta receptor type II (pMSCV-TGFBR-IRES-GFP) or an empty vector (pMSCV-IRES-GF) were kindly provided by Martin Eilers Group, IMT, Marburg, Germany. Cells were incubated with retroviruses containing supernatants for 12 h at 32°C in media containing 4 ?g/ml polybrene. Cells then were expanded at 37°C for an additional 48 h, and GFP-expressing cells were purified by flow cytometry on a FACS Vantage (Becton Dickinson).



All animal experiments were performed by following the guidelines from Administrative Panel on Laboratory Animal Care at Stanford University (protocol 8144). For transplantation experiment cells were washed once with PBS and 10x106 cells/site were injected subcutaneously (s.c.) into the flank of FVB/N syngeneic mice. Tumor growth was measured by using a caliper. To inactivate MYC expression and to assess for tumor regression, the drinking water was supplemented with 200 μg/ml doxycycline. Tumors were harvested untreated or after 1, 3 and 4 days upon doxycycline treatment, respectively. Total RNA was isolated from snap-frozen tumor tissue using the RNeasy Kit including DNase-I digest (Qiagen, Valencia, USA) following the manufacturer's protocol.

ORGANISM(S): Mus musculus

SUBMITTER: Michael Krause 

PROVIDER: E-MEXP-2233 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress