|
| Status |
Public on Jul 15, 2021 |
| Title |
A (hh-) 1 |
| Sample type |
RNA |
| |
|
| Source name |
Non-Hh expressing cells in the anterior (A) compartment of the wing imaginal discs
|
| Organism |
Drosophila melanogaster |
| Characteristics |
tissue: Imaginal discs from wandering third instar larvae
genotype: w; UAS-mCD8-GFP/+; hh-Gal4/+
age: the wandering third instar larval stage
|
| Treatment protocol |
200 pairs of wing imaginal discs were dissected from third instar larvae of the genotypes w; UAS-mCD8GFP/+; hh-Gal4/+ or w; UAS-mCD8GFP/+; ptc-Gal4/+. Discs were stored in Schneider's Drosophila Media plus 10% FBS on ice for less than two hours prior to cell dissociation. Discs were washed twice with 1 ml cell dissociation buffer. Elastase was diluted to 0.4 mg/ml in fresh cell dissociation buffer once discs were ready. Discs were incubated for 20 min at room temperature in 0.4 mg/ml elastase with stirring by a magnetic micro stirring bar. Undissociated tissue was spun out, cell viability was measured using the Beckman Vi-CELL Cell Viability Analyzer (>80%), and cells were immediately isolated using the BD FACSAria II system within the Stanford FACS facility. Dead cells labeled with propidium iodide were excluded during FACS, and purity of sorted cells was greater than 99% by post-sorting FACS analysis.
|
| Growth protocol |
Parental flies, embryos and larvae are grown at 25°C.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from sorted cells (RNeasy, Qiagen), quality was assessed with the Agilent Bioanalyzer 2100 (RIN > 7.0).
|
| Label |
Biotin
|
| Label protocol |
Biotinylated cRNA were prepared according to the standard protocol of the GeneChip® 3’ IVT Express Kit.
|
| |
|
| Hybridization protocol |
Following fragmentation, cRNA were hybridized for 16 hr at 45C on the Affymetrix GeneChip® Drosophila Genome 2.0 Array. GeneChips were washed and stained in the Affymetrix Fluidics Station 450.
|
| Scan protocol |
GeneChips were scanned using the GeneChip® Scanner 3000.
|
| Description |
Gene expression data from A cells
|
| Data processing |
The data were analyzed with the multi-array average (RMA) for background correction followed by quantile normalization. Expression values were log2 transformed.
|
| |
|
| Submission date |
Jul 14, 2021 |
| Last update date |
Jul 15, 2021 |
| Contact name |
Xiaoyan Zheng |
| E-mail(s) |
[email protected]
|
| Phone |
2029944228
|
| Organization name |
George Washington University School of Medicine and Health Sciences
|
| Department |
Anatomy and Cell Biology
|
| Street address |
SEH, Suite 8840, 800 22nd Street NW
|
| City |
Washington |
| State/province |
District of Columbia |
| ZIP/Postal code |
20052 |
| Country |
USA |
| |
|
| Platform ID |
GPL1322 |
| Series (1) |
| GSE180120 |
Genome-wide expression profiling in Drosophila wing imaginal discs |
|
