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. 2013 Aug 30;51(3):301–312. doi: 10.1530/JME-13-0169

Figure 1.

Figure 1

Androgen up-regulates the expression of GNMT mRNA and protein in a time-dependent manner. TaqMan RT-PCR for GNMT (A), PSA (B), NDRG1 (C) and TMPRSS2 (D) was performed from RNA prepared from LNCaP cells cultured in RPMI lacking phenol red supplemented with 10% DSS for 72 h followed by the addition of either ethanol or 1 nM R1881 for 24, 48 and 72 h. Data were normalised to GAPDH levels. The expression of each gene in cells treated with ethanol for 24 h was set to one, with the expression level in the other conditions being shown relative to this. Results are shown as mean values of three replicates with error bars showing s.e.m. (E) LNCaP cells were treated for the indicated times, whole cell lysates were separated by SDS–PAGE and immunoblotted for GNMT and β-actin. The polypeptide corresponding to GNMT is arrowed and molecular weight markers are expressed in kDa. (F) Immunofluorescence staining of GNMT was performed in LNCaP cells treated for 48 h. The GNMT antibody was detected with Alexa Fluor 488-labelled secondary and TO-PRO-3 was used for counterstaining of nuclei. All images were acquired using a Zeiss LSM510 confocal microscope. Scale bar=10 μm.