Figure 5
MCF-7 cells treated with HRG, lactate, CHC and BPTES. The MCF-7 derived knockdown shRNA cells were pretreated with BPTES and CHC for 1 hr. The cells were then stimulated with HRG, lactate or vehicle for 16 hrs. The MCF-7 derived scrambled shRNA cells were pre-treated with BPTES for 1 hr. The cells were then stimulated with HRG, lactate or vehicle for 16 hrs. Samples were harvested with trizol and mRNA was isolated as per manufacturer instructions. qRT-PCR was performed with Sybr Green and ROX as the reference dye to measure the amounts of lactate receptor (GPR81) mRNA. The results were aggregated from the CT data of three samples of different experiments, each performed in technical duplicates. Statistical significance was assessed with the Student’s t-test. Error bars represent the standard deviation. **P ≤ 0.005, ***P ≤ 0.0005.
