Journal: Breast Cancer Research : BCR
Article Title: The sialyl-glycolipid stage-specific embryonic antigen 4 marks a subpopulation of chemotherapy-resistant breast cancer cells with mesenchymal features
Figure Lengend Snippet: Sialyl-glycolipid stage-specific embryonic antigen 4 (SSEA4) expression is regulated by CMP- N -acetylneuraminate-β-galactosamide-α-2,3-sialyltransferase 2 ( ST3GAL2 ). a Small interfering RNA (siRNA)–mediated knockdown of ST3GAL2 and ST3GAL3 significantly reduced the expression of their respective target messenger RNAs (mRNAs), but not the housekeeping gene GAPDH , as measured by quantitative real-time polymerase chain reaction. Each bar represents the expression intensity normalized to the Lipofectamine reagent (Life Technologies, Carlsbad, CA, USA)-only control. b Knockdown of ST3GAL2 significantly reduced the expression of SSEA4, whereas targeting of CD133 or the close paralog ST3GAL3 did not result in a significant change of SSEA4 expression. Knockdown of CD133 expression by the respective siRNA was used as a positive control of direct targeting. Each bar represents the mean fluorescence intensity (MFI) normalized to the Lipofectamine-only control. c Expression of ST3GAL2 mRNA was measured by two Affymetrix® (Affymetrix, Santa Clara, CA, USA) probes, 217650_x_at and 229336_at, in nine tumor models and plotted against the frequency of SSEA4-positive cells as measured by flow cytometry in the respective models, which showed a significant positive correlation. *** p
Article Snippet: Microarray hybridization and data analysis Messenger RNA (mRNA) and microRNA (miRNA) expression profiling was performed using Agilent microarrays (Agilent Technologies, Santa Clara, CA, USA) according to the manufacturer’s instructions.
Techniques: Expressing, Small Interfering RNA, Real-time Polymerase Chain Reaction, Positive Control, Fluorescence, Flow Cytometry, Cytometry