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. 2001 Mar 13;98(7):3837–3842. doi: 10.1073/pnas.061358098

Figure 1.

Figure 1

Endogenous menin expression is suppressed in GH4C1 cells stably transfected with antisense menin cDNA. (a) Scheme for construction of the antisense menin vector. MCS, multiple cloning site; Neomycin, neomycin resistance gene; PCMV, human cytomegalovirus immediate-early promoter/enhancer; E, EcoRI; S, SacI. The asterisk indicates the stop codon at position 611 in the menin cDNA. (b) Menin expression in empty vector-transfected (V)- and antisense menin (AS)-transfected GH4C1 cells was assessed by immunoblotting. Equivalent amounts of total protein were loaded in each lane. (c) Human menin antisense transcripts were assessed by reverse transcription–PCR. GAPDH, glyceraldehyde-3-phosphate dehydrogenase. (d and e) TGF-β stimulates menin expression in GH4C1 cells. Serum-starved cells were cultured (d) in the presence of various concentrations of TGF-β for 1 h, (e) in the presence of 100 pM TGF-β for the indicated times, or (f) in the presence of 100 pM TGF-β for 1 h. Total cell lysates (30 μg) were separated by SDS/PAGE, immunoblotted, and revealed with anti-menin antibody. As a control for protein loading, membranes were probed with an anti-Stat3 antibody.