Figure 2
From: Integrated analysis of mRNA and miRNA expression in HeLa cells expressing low levels of Nucleolin
Transcriptomic analysis of nucleolin depleted cells. (A) HeLa cells were treated with control or NCL siRNA as previously described27, 28 and after 5 days, protein extracts and total RNA were prepared. NCL protein expression was assessed by western blot and normalized by comparison with β-actin (left panel) while NCL mRNA level was measured by RT-qPCR (right panel). (B) Lists of the 20 most down or up- regulated genes and their fold change (FC) and p-values with top 20 pathways and GOBP terms enriched with least FDR-q value. (C) RT-qPCR validation of genes that were found either up-, down- or unchanged in the microarray analysis. NCL was included in the analysis, together with some genes involved in cell cycle regulation and other randomly chosen genes. (D) Western blot analysis with total extract of the expression of CCNB1 protein upon the depletion of NCL. As CCNB1 was strongly down-regulated and may be important for the arrest of HeLa cells in G2/M phase upon NCL depletion, we choose this protein to check for the correspondence between RNA level (Affymetrix microarrays-RT-qPCR) and protein levels.
