HUVECs cells were grown in 10 cm dish plate and starved overnight in 1% medium 200 before the experiment. Cells were treated when they are at 70-90% confluence with 100 nM human insulin or 5uM SB431542. Cells were collected after 90 min or 6 h treatment. Cells were washed in ice cold PBS twice before collected and process for RNA purification using RNAeasy Plus (QIAGEN) with DNAseI treatment step.  10% FBS Medium 200 with LSGS Supplement from Invitrogen Pretreatment with SB431542 for 45 min. RNA extraction was performed using RNAeasy Plus with DNAseI treatment step.  Library was produced using NUGEN Ovation human FFPE rnaseq library system
Experimental Factor: time
1.5
Experimental Factor: stimulus
none