Figure 5. Plac8 expression during cadmium-induced RWPE-1 transformation.

(A) RWPE-1 cells treated with cadmium for 0 (untreated), 2 and 4 months and non-treated CTPE cells were grown in soft agar for 10–15 days. Colonies were stained with crystal violet and counted. (B) Levels of Plac8 expression in RWPE-1 cells as a function of cadmium exposure time (months, mo) and non-treated CTPE cells were measured in whole cell lysates by Western blot analysis; β-actin was used as a loading control. (C) Total RNA was extracted and qRT-PCR was performed to examine steady state levels of Plac8 mRNA expression in RWPE-1 exposed to cadmium for 0 (untreated) 2 and 4 months and CTPE cells. (D) Intracellular location of Plac8 was determined by immunohistochemistry in cadmium-treated and non-treated RWPE-1 and CTPE cells. Student’s t-tests were used to calculate statistically significance differences between the various treatment groups. *p < 0.05, **p < 0.01, ****p < 0.0001