LARGE2 mRNA expression correlates with hypoglycosylation of αDG.
A, mRNA analysis by qRT-PCR shows dramatically reduced LARGE2 expression, compared with other genes known to influence αDG glycosylation, in TEM4–18 cells compared with PC-3E+ cells. B, LARGE2 mRNA is detectable in the prostate cancer cell line panel and is greatly reduced in TEM4–18 and GS689.Li cells. All LARGE2 samples validated with two probe sets on two separate RNA isolations. Numbers on top of bars are Ct values: GAPDH/gene of interest. C, immunofluorescence staining shows αDG glycosylation (IIH6) is present in a WT mouse prostate and the myodystrophy mouse (Largemyd, MDC1D) prostate (arrows). Staining is not present in a prostate-specific DG knock-out mouse. Muscle staining (asterisks) indicates tissue-specific deletion. Bar, 50 μm.