A TRPC1 protein-dependent pathway regulates osteoclast formation and function - PubMed (original) (raw)

Generation of _I_SOC by TRPC1α and amplification of _I_CRAC by TRPC1ϵ. A, time course and I–V curve (taken at 200 s) of BAPTA-induced whole cell currents in cells transfected with STIM1 (1.6 μg), Orai1 (1 μg), and TRPC1α (1 μg), (n = 9). pF, picofarad. B, time course and I–V curve (taken at 200 s) of BAPTA-induced whole cell currents in cells transfected with STIM1 (1.6 μg), Orai1 (1 μg), and TRPC1α-ΔNTx (1 μg) (n = 10). C, time course and I–V curve (taken at 200 s) of BAPTA-induced whole cell currents in cells transfected with STIM1 (1.6 μg), Orai1 (1 μg), and TRPC1ϵ-ΔNTx (1 μg) (n = 14). D, time course and I–V curve (taken at 200 s) of BAPTA-induced whole cell currents in cells transfected with STIM1 (1.6 μg), Orai1 (1 μg), and TRPC1ϵ (1 μg) (_OST_ϵ, red, n = 11) or STIM1 (1.6 μg) and Orai1 (1 μg) (OS, blue, n = 8). E, expression levels of Orai1, STIM1, TRPC1, or CD8α in transfected HEK293T cells. Cell lysates transfected with GFP (negative control, lane 1), Orai1, STIM1, and CD8α (lane 2), Orai1, STIM1, TRPC1α-ΔNTx, and CD8α (lane 3), Orai1, STIM1, TRPC1α, and CD8α (lane 4), Orai1, STIM1, TRPC1ϵ-ΔNTx, and CD8α (lane 5), or Orai1, STIM1, TRPC1ϵ, and CD8α (lane 6) were immunoblotted with a rabbit polyclonal antibody to Orai1 (1:5000, Sigma), mouse monoclonal antibody to STIM1 (1:1000, Cell Signaling), mouse monoclonal to TRPC1 (1F1, 1:300), or rabbit polyclonal to CD8α (1:500, Santa Cruz Biotechnology). Asterisks indicate nonspecific bands. F, effects of Na+ to _N_-methyl-

-glucamine substitution on whole cell currents induced by BAPTA (10 m

in pipette) in HEK293 cells transiently transfected with Orai1 and STIM1 (OS, black, (n = 5) or Orai1, STIM1, and TRPC1ϵ (O_ST_ϵ, red, n = 8). G, effects of Ca2+ to Ba2+ substitution on whole cell currents induced by BAPTA (10 m

in pipette) in HEK293 cells transiently transfected with Orai1 and STIM1 (OS, black, n = 7) or Orai1, STIM1, and TRPC1ϵ (OSTϵ, red, n = 8).