Fig. 2.

Differentiated visceral adipocytes also exhibit increase in SOCE. (A,B) Western blots (A) of TRPC1, ORAI1, Stim1, FAPB4 and perilipin, and actin protein expression, protein level quantification normalized to actin provided as bar graph (B), of VAT SVF and differentiated VAT (n=3). (C–H) Application of 1 μM Tg induced inward currents at −80 mV in control and SKF treated VAT-SVF cells (C) and differentiated VAT (F). Respective I–V curves are shown for VAT SVF (D) and differentiated VAT (G). Quantitation (n=6 recordings) of current intensity at −80 mV for VAT SVF (E) and differentiated VAT (H). (I,J) Western blot (I) and protein level quantification (J, normalized to actin and presented as fold change relative to control untreated cells) of PPARγ, FAPB4 and perilipin protein expression of differentiated VAT treated with SKF (n=3–4). (K) Quantification of eluted Oil-Red-O stain at 492 nm of differentiated VAT in the presence of SKF (10 µM) for 7 days (n=3). (L–O) Fura-2 fluorescence traces of transient increase in [Ca²⁺]_i after addition of 1 μM Tg and 1 mM Ca²⁺ to VAT SVF cells (L) and differentiated VAT (N) pretreated with 10 μM SKF for 15 min. Quantification of Tg and Ca²⁺ peaks for VAT SVF cells (M) and differentiated VAT (O). Each bar gives the mean±s.e.m. of 40–60 cells in three separate experiments. Graphs are mean±s.e.m. *P<0.05, **P<0.01, ****P<0.0001 using one-way ANOVA.