NextSeq 500 sequencing; RNA-seq of ZFP30 knock-down and knock-out in mouse 3T3-L1 and IBA cells during adipogenic differentiation
RNA-seq of ZFP30 knock-down and knock-out in mouse 3T3-L1 and IBA cells during adipogenic differentiation
3T3-L1 or IBA cells were induced to differentiation at day 0 (2 days post-confluence) by adding the induction medium, which is complete culture medium supplemented with the MDI cocktail (1μM dexameth¬asone, 0.5 mM 3-isobutyl-1-methylxanthine, and 167 nM insulin, all from Sigma, Saint-Louis, MO). At day 2, induction medium was removed and maintainance medium (complete culture medium supplemented with 167 nM insulin) was added. At day 4, the maintenance medium was removed and complete culture medium was added. The cells were harvested for staining or RNA extraction at day 6.  RNA extractions were performed using the Direct-zol RNA Miniprep kit (R2052, Zymo Research). Potential genomic DNA contamination was eliminated by Dnase I digestion. The RNA quality and concentration was checked by Nanodrop spectrophotometer (Thermo Scientific).  RNA extractions were performed using the Direct-zol RNA Miniprep kit (R2052, Zymo Research). Potential genomic DNA contamination was eliminated by Dnase I digestion. The RNA quality and concentration was checked by Nanodrop spectrophotometer (Thermo Scientific).
Experimental Factor: cell line
IBA
Experimental Factor: genotype
Zfp30 knockout
Experimental Factor: time
0