|Application:||Gene expression microarray analysis|
|Number of samples:||6|
|Release date:||Jun 13 2018|
|Last update date:||Jun 13 2018|
|Diseases:||Breast Neoplasms, Neoplasms, Germ Cell and Embryonal, Neoplasms, Glandular and Epithelial|
|Dataset link||ERK3 is essential for establishment of epithelial architecture [ERK3 KD vs. TFAP2A KD]|
Control MO (80 ng), ERK3 MO1/2 (40 ng each of MO1 and MO2), or TFAP2A MO1/2 (40 ng each of MO1 and MO2) were injected into the animal regions of all blastomeres at the 4-cell stage. The animal caps were dissected from the injected embryos at stage 9, cultured alone, and harvested at stage 15. Two independent replicates were prepared. Total RNA was extracted using TRIzol reagent. The quality of the total RNA was assessed using an Agilent 2100 BioAnalyzer. cDNA synthesis and transcriptional amplification were performed using 250 ng of total RNA with the GeneChip 3’ IVT PLUS Reagent Kit (Affymetrix, #902415). Fragmented and biotin-labeled cDNA targets were hybridized to the GeneChip Xenopus laevis Genome 2.0 Array (Affymetrix) according to the manufacturer's protocol. Hybridized arrays were scanned using an Affymetrix GeneChip Scanner. Scanned chip images were analyzed with GeneChip Operating Software v.1.4 (GCOS). The probe set signal intensities in the raw data (CEL files) were normalized using a robust multiarray average (RMA) algorithm and Expression Console software.