Lipid droplet quantification (ArrayScan)

Cellular LDs were quantified adapting the Cellomics™ assay described previously [19] applying the Thermo Scientific™ ArrayScan™ VTI High Content Reader (Thermo Fisher Scientific, Waltham, MA, US). Briefly, differentiated adipocytes (day 14) were fixed, stained with the fluorescent dyes Hoechst 33342 (nuclei) and BODIPY® 493/503 (LDs; Life Technologies) followed by quantification of accumulated LDs with the SpotDetector® V2 algorithm. Adjusted protocol parameters are listed in Additional file 3: Table S2. For analysis 100 fields per well were scanned and data of each channel were reported on a “per field” basis. The nuclei related features Spot Count (= LD-number), Spot Total Area (= LD-area) and Spot Total Intensity (= LD-intensity), describing the differentiation status of treated adipocytes, were calculated as percent of Diff CTRL per plate.