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. 2001 Apr;12(4):831–845. doi: 10.1091/mbc.12.4.831

Figure 1.

Figure 1

Expression of dominant negative Cx in parental and transfected bEnd.3 cells. (A) mRNA isolated from parental and transfected cells was amplified in a RT-PCR reaction using primer pairs specific for Cx43 (lanes 1–5), 3243H7 (lanes 6–8), or Cx43βGal (lanes 9–11). An aliquot of the PCR amplification reaction was electrophoresed in a 2% agarose gel. Lanes 1, 6, and 9: parental bEnd.3 cells; lanes 2 and 7: bEnd.3/3243H7 cells clone B3; lanes 3 and 8: bEnd.3/3243H7 cells clone B5; lanes 4 and 10: bEnd.3/Cx43βGal cells clone D1; lanes 5 and 11: bEnd.3/Cx43βGal cells clone D2; and lane m:100-bp ladder. (B–E) Subconfluent cultures of bEnd.3/3243H7 and bEnd.3/Cx43βGal cells were incubated with anti-HA and anti-βGalactosidase antibodies, respectively, as well as with anti-Cx43 antibodies. In bEnd.3/3243H7 cells, the chimeric protein (B) was detected predominantly in the perinuclear region, although some diffuse fluorescence was also present throughout the cytoplasm. Note the absence of immunostaining at the sites of cell-to-cell contact. In bEnd.3/Cx43-βGal cells (D), the fusion protein was detected as punctate spots in regions of cell-to-cell contact and in the perinuclear region of the cytoplasm. Similar labeling patterns were observed using Cx43 antibodies (C and E). Bar, 30 μm.