FIGURE 3.

GOLPH3 knockdown reduces functional glycosylation of α-dystroglycan. A, HEK293 cells were transfected with POMGnT1 siRNA, followed by FACS analysis with IIH6 antibody. B, HEK293 cells were transfected with GOLPH3 siRNA, followed by FACS analysis with IIH6 antibody. Control (Ctrl) siRNA indicates non-targeting siRNA control. NC, negative control without the primary antibody. Cell surface α-dystroglycan produced in cells treated with POMGnT1 and GOLPH3 siRNA displayed reduced IIH6 immunoreactivity. The relative POMGnT1 (C) and GOLPH3 mRNA levels (D) following transfection with the corresponding siRNAs were quantified by qRT-PCR. Bar charts summarize data obtained from three replicates and was normalized against endogenous GAPDH. Error bars indicate S.E. or S.D. A significant reduction in the expression of both genes was observed.