R31C GNRH1 Mutation and Congenital Hypogonadotropic Hypogonadism
Figure 7
Effects of treatments on lhb transcription and LH secretion.
(A) LbetaT2 cells were starved overnight in serum-free DMEM and treated either by 10 nM WT, R31C, 10 nM WT +10 nM R31C peptides or vehicle. After five hours treatment cells were lysed for RNA extraction (Trizol®). Mouse lhb levels were analyzed by quantitative real time RT-qPCR. Values are expressed as individual ratios on 18S and represented as fold induction on vehicle as means ± SEM, **p<0.01. This experience was conducted n = 7 times in duplicates. (B) LH concentrations from LbetaT2 cells culture media. Cells were starved overnight in serum-free DMEM and incubated with 10 nM WT, 10 nM R31C, 10 nM WT +10 nM R31C, or vehicle for 5 hours. LH levels were measured by a combined rat/mouse RIA. Values are expressed as individual values and as means ± SEM, *p<0.05, ***p<0.001. The experiment was conducted four times and each sample was assayed in duplicates.