Figure 1
From: Regulation of cell proliferation and cell density by the inorganic phosphate transporter PiT1
Knock-down of PiT1 in MC3T3-E1 cells decreases overall proliferation and cell density. A and B) qRT-PCR analyses. MC3T3-E1-PiT1shRNA (PiT1shRNA) and MC3T3-E1-LMP (Control) cells were seeded at 20,000 cells/cm2 and after 2 days in culture, the mPiT1 (A) and mPiT2 (B) mRNA levels were analyzed by qRT-PCR. Each column represents cell lysates from three wells and triplicate qRT-PCR analyses of each cell lysate. The mPiT1 and mPiT2 mRNA levels are standardized to B2M mRNA levels. Data are means ± standard deviation (SD).*indicates statistically significantly different from control cells, p < 0.05. C) Pi-transport assay. MC3T3-E1-PiT1shRNA (PiT1shRNA) and MC3T3-E1-LMP (Control) cells were seeded at 50,000 cells/cm2 in 4-well plates. Two days after, 32Pi import in Pi-free medium (with 5 μM 32Pi only) was analyzed over 5 minutes. The results are shown as mean 32Pi import per mg protein per hour of 4 wells ± SD. D) Cell counts. MC3T3-E1-PiT1shRNA (PiT1shRNA) and MC3T3-E1- LMP (Control) cells were seeded at 20,000 cells/cm2 in 4-well plates and counted at the indicated days. The results are shown as mean cell number per cm2 of 4 wells ± SD.* indicates statistically significantly different from control cells at the same day, p < 0.05.