Figure 2

Effects of RNA interference-mediated knock-down on lipolysis and lipids in human adipose-derived stem cells. hASCs were transfected with control siRNA oligonucleotide (siNegC) or indicated siRNAs targeting specific genes. (a,b) Expression of ZNF436, WARS2, TBX15, INVS and STX17 was knocked down using siRNA in hASCs 24 h prior induction of differentiation and followed until differentiation day 9, upon which accumulated glycerol in medium was measured to assess spontaneous lipolysis (a) and accumulation of neutral lipids was evaluated by lipid staining (b). (c,e) Expression of PCK1, EBPL, NID2, GGA3, MRPS7, NUP85, GRB2 and PEMT was knocked down using siRNA in hASCs 24 h prior induction of differentiation and followed until differentiation day 9, upon which glycerol in medium was measured to assess spontaneous lipolysis (c) and accumulation of neutral lipids was evaluated in the cells (e). Results are based on four biological/independent experiments. Results were analyzed using t-test and presented in fold change ± SD relative to negative control Neg C. ***p < 0.005, **p < 0.01, *p < 0.05. (d) Expression of NUP85 was knocked down using siRNA in hASCs 24 h before induction of adipogenenic differentiation of hASCs. At day 9 of differentiation cells were incubated with isoprenaline (ISO) and the effects of knock-down on lipolysis were evaluated. Results are based on four biological/independent experiments. Results were analyzed using t-test and are presented as fold change ± SD relative to 0 M isoprenaline (ISO) of respective condition (siNegC or siNUP85). ***p < 0.005, **p < 0.01.