Journal: The EMBO Journal
Article Title: Asf1b, the necessary Asf1 isoform for proliferation, is predictive of outcome in breast cancer
doi: 10.1038/emboj.2010.335
Figure Lengend Snippet: Distinct effects of Asf1a and Asf1b depletions. (A) (Left panel) Western blot analysis of total extracts from human U-2-OS cells showing the specific depletion of Asf1a, Asf1b or Asf1(a+b) for 48 h by siRNA treatment. Increasing amounts (X) of total cell extracts are loaded and α-Tubulin serves as a loading control. A mix of the specific Asf1 antibodies reveals Asf1a and Asf1b. M: molecular weight marker. (Right panel) Flow cytometry analysis of the cell cycle distribution of the cells shown in the left panel. (B) mRNA extracted from human U-2-OS cells treated as in A were hybridized to Affymetrix HG-U133-Plus2 oligonucleotide microarrays. mRNA expression levels of Asf1a and Asf1b obtained from the Affymetrix hybridization are expressed as a log2 (fold change) relative to the control siRNA depletion. Error bars represents data from two independent experiments. (C) Venn diagram showing the overlap between the significantly (P<0.05) differentially expressed genes determined in each siRNA condition indicated (siAsf1a, siAsf1b and siAsf1(a+b)) versus the control siRNA. Numbers indicate the quantity of genes overlapping between two conditions. (D) Gene ontology (GO) analysis of differentially expressed genes after the specific knockdown of Asf1a or Asf1b or Asf1(a+b) (see Supplementary data for details). Histogram bars represent the −log(P-value) for each significant GO term. Terms for which the test resulted in a −log(P-value) equal to 3.5 (P-value <5.10−4) as shown by the dashed line were considered to be significantly overrepresented for the given list. Numbers in brackets indicate the number of significant genes found in the list of differentially expressed genes for the given GO category. Arrows indicate if the genes are upregulated or downregulated in the indicated siRNA relative to the control siRNA.
Article Snippet: Transcriptomic data analysis In two independent experiments, we prepared mRNAs using U-2-OS cells treated with control, Asf1a, Asf1b or Asf1(a+b) siRNAs for 48 h, and hybridized them on Affymetrix HG-U133-Plus2 oligonucleotide microarrays.
Techniques: Western Blot, Molecular Weight, Marker, Flow Cytometry, Expressing, Hybridization
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