Endogenous expression of TRPC3 in human hematopoietic cells. Western
blotting was performed on lysates from UT-7 and TF-1 Epo-responsive cell
lines, from CD34+ cells and from day 10 and 14 BFU-E-derived
erythroblasts. Equivalent amounts of protein were loaded in each lane.
A, immunoblotting with anti-TRPC3 antibody demonstrated increased
expression of TRPC3 in primary human erythroid cells during erythroid
differentiation. Blots were probed with anti-actin antibody to compare loading
of lanes. Representative results of four experiments are shown.
B, densitometry was used to quantitate TRPC3 and actin bands from
four experiments of lysates from CD34+ cells and day 10 and 14
BFU-E-derived erythroblasts. The TRPC3/actin ratio was calculated and
normalized to CD34+ cells to allow comparison between experiments,
and the mean normalized ratio ± S.E. was determined. TRPC3 expression
was significantly less in CD34+ cells than in day 10 erythroblasts
(p < 0.02). C, immunoblotting with anti-Epo-R antibody
demonstrated greatest Epo-R expression in day 10 primary erythroblasts.
Representative results of three experiments are shown.