Transcription profiling by high throughput sequencing of HNRNPC knockdown and control HeLa cells
Transcription profiling by high throughput sequencing of HNRNPC knockdown and control HeLa cells
HeLa cells were grown in Dulbeccos modified Eagle medium (DMEM) supplemented with 10% Fetal Bovine Serum and 1% penicillin-streptomycin and cultured at 37C with 5% CO2. For the knockdown of HNRNPC, HeLa cells were plated at concentration of 105 cells/well in a 6-well cluster plate in 2 ml of growth medium. 24 h after the plating (when confluency reached around 20%), cultures were independently transfected using two different hnRNP C Stealth Select RNAi siRNAs (KD1 and KD2 refer to siRNAs HSS179304 and HSS179305 from Invitrogen, respectively) at a final concentration of 5 nM. Both siRNAs were transfected using Lipofectamine RNAiMAX (Invitrogen) according to the manufacturers instructions. Using the same procedure, we used Stealth RNAi siRNA Negative Control (Invitrogen) for the control experiments. Library preparation was preformed according to mRNA Sequencing Sample Preparation Guide (Illumina, part # 1004898 REV. D). Reagents were taken from the Illumina sample preparation kit (Illumina, CAT # RS-930-1001).
Experimental Factor: RNAI
KD1