FIGURE 5:

Perilipin S497 and S522 phosphorylation upon isoproterenol stimulation in OPA1 knockdown and control conditions. hASC-derived adipocytes were transfected with OPA1 siRNA or scrambled control 3 wk after initiation of differentiation and stimulated with 0–100 nM isoproterenol for 3 min before immunoblotting for the presence of OPA1, perilipin 1 pS522 and pS497, perilipin 1, mitofilin, and actin (A). Densitometric analysis of perilipin 1 phosphorylation levels at S522 and S497 as in A normalized against actin (mean ± SEM from 3 donors) under scrambled control (▪) or siOPA1 (▴) conditions (B). Immunofluorescence staining for perilipin 1 pS522 or pS497 (green) and DAPI (blue) in hASC-derived adipocytes transfected with OPA1 siRNA or scrambled control, ± 10 nM of isoproterenol for 3 min (C). Scale bars: 10 µm. Statistical analysis of experiments as in C; ***p < 0.0005, *p < 0.05 (D). One hundred cells were scored from each condition and for every donor (mean ± SEM, n = 3 donors).