Figure 5.

Effects of GTRAP3-18 antisense oligonucleotides in MeβCD-treated CGNs. A, After CGNs had been treated with 250 μm MeβCD, CGNs were transiently cotransfected with pGFP and GTRAP3-18 sense or antisense oligonucleotides. Two days later, immunocytochemistry using anti-GTRAP3-18 antibody (DsRed) was performed. GFP-positive cells were considered to be oligonucleotide-transfected cells. B, After CGNs had been treated with 250 μm MeβCD, CGNs were transiently cotransfected with pDsRed and GTRAP3-18 sense or antisense oligonucleotides. Two days later, the cells were stained with CMFDA (CMF; green). DsRed-positive cells were considered to be oligonucleotide-transfected cells. C, Fluorescence intensity changes on both MeβCD-treated and oligonucleotide-transfected cells (GFP-positive or DsRed-positive cells) were quantified in comparison with the intensity of nontreated, nontransfected cells, the value for which was taken as 0%. The fluorescence intensities of GFP- and DsRed-positive cells were quantified in two independent experiments with triplicate culture wells. The average number of GTP- or DsRed-positive cells was 5–10. *p < 0.05 compared with control; **p < 0.05 compared with both MeβCD-treated and GTRAP3-18 sense oligonucleotide-transfected cells.