Figure 4
Mitochondrial Ca2+ intake through the Ca2+ uniporter is reduced in HT1080/BI-1 cells.
(A) HT1080/Neo and HT1080/BI-1 were loaded with Rhod II-AM and subsequently treated with 5 μM thapsigargin, 5 μg/ml tunicamycin, or 1 μM brefeldin A in the presence or absence of 1 μM RU360. Rhod II fluorescence was analyzed. (B) HT1080/Neo and HT1080/BI-1 were transfected with erAEQ or loaded with Rhod II and treated with 5 μM thapsigargin, 5 μg/ml tunicamycin, or 1 μg/ml brefeldin A in the presence or absence of 1 μM RU360. The intensity of erAEQ luminescence or Rhod II fluorescence was quantified. *, p < 0.05 versus thapsigargin-treated HT1080/Neo; #, p < 0.05 versus tunicamycin-treated HT1080/Neo; &, p < 0.05 versus brefeldin A-treated HT1080/Neo. (C) Mitochondrial 45Ca2+ intake was analyzed in HT1080/Neo and HT1080/BI-1 in the presence or absence of RU360. *, p < 0.05 versus HT1080/Neo without RU360. (D) Viability of HT1080/Neo and HT1080/BI-1 cells after treatment with 5 μM thapsigargin in the presence or absence of RU360 was determined by trypan blue staining. *, p < 0.05 versus thapsigargin-treated HT1080/Neo Tg, thapsigargin; Tu, tunicamycin; Bre, brefeldin; Con, Control; Neo, neomycin-resistant pcDNA3-transfected HT1080 cells (HT1080/Neo); BI-1, HA-BI-1-pcDNA3-transfected HT1080 cells (HT1080/BI-1).
