Figure 3
D 2 in human GCs. A) Result of cDNA array, showing evidence for expression of D2. B) Ethidium bromide-stained agarose gel showing RT-PCR products which identify D2L (261 bp) and D2S (173 bp). Note that D2L is consistently expressed, while D2S expression is not depending on age or treatment with hCG (day 4 of culture, n = 4/each). C) Western blot analysis of D2 in cultured human GCs. Cells were treated without or with hCG for 4 days and 15 μg of total protein was analyzed. Note that the treatment has no effect on the overall levels of D2 protein, which can be readily detected as a double band of about 48 and 51 kDa, representing D2S and D2L, respectively. Subsequent detection of beta actin in the same blots was also performed to normalize the density of the bands. Data shown are from one of three independent experiments yielding similar results.