|
| Status |
Public on May 15, 2023 |
| Title |
25mM ALA treatment, harvest at 24h Replicate 2 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
HepG2 cell line (code 0103)
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: HepG2
time: 24 h
treatment: ALA 25 mM
|
| Treatment protocol |
After reaching 80% confluence, the cells were treated or not with ALA 2.5 mM or 25 mM for 2 h or 24 h (identified as: Control, ‘2.5mM-2h’, ‘2.5mM-24h’, ‘25mM-2h’, ‘25mM-24h’)
|
| Growth protocol |
The HepG2 cell line (code 0103), from Rio de Janeiro Cell Bank (BR), was cultivated in DMEM-low glucose with 100 IU/mL penicillin and 0.1 mg/mL streptomycin (Thermo Fisher, USA), supplemented with 10% fetal bovine serum (FBS) (Cultilab, BR), and maintained at 37° C in 5% CO2 atmosphere.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted using the RNAspin Mini RNA isolation kit (GE Healthcare, USA)
|
| Label |
Cy5
|
| Label protocol |
Double-stranded cDNA was generated from 100 ng total RNA using the Low Input Quick Amp Labeling Kit, two-color with Cyanine 3-CTP or Cyanine 5-CTP.
|
| |
|
| Channel 2 |
| Source name |
Control
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: HepG2
treatment: ALA 0 mM
time: 24 h
|
| Treatment protocol |
After reaching 80% confluence, the cells were treated or not with ALA 2.5 mM or 25 mM for 2 h or 24 h (identified as: Control, ‘2.5mM-2h’, ‘2.5mM-24h’, ‘25mM-2h’, ‘25mM-24h’)
|
| Growth protocol |
The HepG2 cell line (code 0103), from Rio de Janeiro Cell Bank (BR), was cultivated in DMEM-low glucose with 100 IU/mL penicillin and 0.1 mg/mL streptomycin (Thermo Fisher, USA), supplemented with 10% fetal bovine serum (FBS) (Cultilab, BR), and maintained at 37° C in 5% CO2 atmosphere.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted using the RNAspin Mini RNA isolation kit (GE Healthcare, USA)
|
| Label |
Cy3
|
| Label protocol |
Double-stranded cDNA was generated from 100 ng total RNA using the Low Input Quick Amp Labeling Kit, two-color with Cyanine 3-CTP or Cyanine 5-CTP.
|
| |
|
| |
| Hybridization protocol |
RNA Spike-In Kit, two-color, was used as a positive hybridization control.
|
| Scan protocol |
Scanned on an Microarray Agilent SureScan scanner using AgilentG3_GX_2color protocol.
Images were quantified Agilent Feture Extraction Software (version A.10.7.1.1).
|
| Description |
Biological replicate 2 of 4. treated with 25 mM ALA and harvested after 24 h.
|
| Data processing |
Agilent Feature Extraction Software was used for background subtraction and LOWESS normalization.
|
| |
|
| Submission date |
May 20, 2022 |
| Last update date |
May 15, 2023 |
| Contact name |
Janice Onuki |
| E-mail(s) |
janice.onuki@butantan.gov.br
|
| Phone |
+55 11 97304-0447
|
| Organization name |
Butantan Institute
|
| Street address |
Av. Vital Brasil, 1500 - Butantã
|
| City |
São Paulo |
| State/province |
SP |
| ZIP/Postal code |
05503-900 |
| Country |
Brazil |
| |
|
| Platform ID |
GPL20844 |
| Series (1) |
| GSE203437 |
Transcriptome profile analysis reveals putative molecular mechanisms of 5-aminolevulinic acid toxicity |
|
