Silica gel on tlc al foils

Cells were seeded and differentiated in 6‐well plate. On day 7 of differentiation, cells were incubated with 1 µM Bodipy 558/568 C12 and DMSO, 100 nM UK5099 or 1 µM norepinephrine for 4 and 24 h. Thin‐layer chromatography of intracellular lipids and extracellular lipids was performed as previously described (Rambold et al, 2015) with minor modifications. Intracellular lipids and lipids from media were extracted in 500 µl chloroform. Chloroform was evaporated using the Genevac EZ‐2 Plus Evaporating System (Genevac, Ipswich, United Kingdom). Lipids were then dissolved in 15 µl chloroform, and 1 µl was spotted on a TLC plate (Silica gel on TLC Al foils, Sigma‐Aldrich, St. Louis, MO). Lipids were resolved based on polarity in a developer solution containing ethylacetate and cyclohexane in a 2.5:1 ratio. TLC plates were imaged on the ChemiDoc MP imaging system (Bio‐Rad Laboratories, Hercules, CA). Band densitometry was quantified using FIJI (ImageJ, NIH).

Solvents (ethyl levulinate, methyl levulinate, 2-MeTHF, and dichloromethane), substrates (1–19), nucleophiles (a–h), and triethylamine (Et₃N) were purchased from Sigma-Aldrich (St. Louis, MO, USA), and they were used without any further purification. The Pd(OAc)₂ and the ligands (PPh₃ and XantPhos) were also purchased from Sigma-Aldrich. TLC plates (silica gel on TLC Al foils with fluorescence indicators of 254 nm) and the silica gel (high-purity grade, average pore size 60 Å (52–73 Å), 70–230 mesh) were purchased from Sigma-Aldrich (St. Louis, MO, USA).