The Effect of OPA1 on Mitochondrial Ca2+ Signaling (original) (raw)
Figure 2
Effect of OPA1 or Mfn1 silencing on Ca2+ signaling in intact H295R cells.
The cells were transfected with control RNA or siRNA and 4mt-D2-cpV on the day following plating (day 2)and once again with control RNA or siRNA on day 3. On day 5, after preloading with Fura-2 AM, the cells were stimulated with 25 mM K+. Changes in cytosolic [Ca2+] ([Ca2+]c) were monitored by measuring Fura-2 excitation ratio while [Ca2+]m was indicated by the FRET ratio of 4mt-D2-cpV. Both ratios are normalized to those obtained in the control period. Representative cytosolic Ca2+ signals (A) and mitochondrial Ca2+ uptake curves (B) are shown for cells transfected with control RNA, Mfn1 or OPA1 siRNA. C: peak [Ca2+]c, D: peak [Ca2+]m E: [Ca2+]m response normalized to peak [Ca2+]c (Δ[Ca2+]m/[Ca2+]c) indicating mitochondrial responsiveness; F: the slope of [Ca2+]m rise related to peak [Ca2+]c (cells not displaying a mitochondrial Ca2+ response were omitted from this statistics). Data are shown for control (C), Mfn1 siRNA (M) or OPA1 siRNA-transfected (O) groups. Results represent mean + SEM, the number of observations is shown within the columns.