External factors
| External factors: |
Etoposide |
| Category: |
Chemical compounds |
| Phenotype: |
Embryonal carcinoma |
| Experiment: |
Flow cytometry//Immunofluorescence//Cell morphological analysis |
| Description: |
G2M arrest and nuclear swelling were evident on day 3 after ETO treatment, with restoration of normal cell cycle and nuclear size 4 d later. Concurrent nuclear area assessments and DNA content measurements demonstrate that the nuclei of ETO treated cells increased in size irrespective of the stage of the cell cycle, but was most evident in G2M and polyploid cells. Increased nuclear area and DNA content were also accompanied by an increase in cellular granularity, as determined by an increase in side scatter detected by flow cytometry analysis, and autophagy (see below). The majority of cells displayed flattened morphology, but only a proportion displayed p16Ink4a nuclear positivity. Furthermore, p16Ink4a expression was largely confined to the cytoplasm . |
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Regulatory relationship
| Target gene:
|
OCT4A//P21//P53//P16 |
| R-EF-Target gene:
|
Activation//Upregulation//Upregulation//-- |
| Official symbol(s): |
POU5F1//P21//P53//P16 |
| Target gene experiment: |
Flow cytometry//Western blot |
| Target gene description: |
Both the expression and variation of expression of OCT4A and p21Cip1 increased on day 3 after ETO treatment; and increased further on day 5 and a high number of double-positive cells confirmed.Immunoblotting confirmed that p53 increased in response to ETO from day 1, resulting in the induction of p21Cip1, which reached its maximum on day 5. In ETO-treated cells (day 4), the cytoplasm was highly enriched with LAMP2, while p16ink4a-positive aggresomes (larger and more numerous than in control) were sequestered in autophago-lysosomes. |
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Aging network
Annotation:
The green line represents Upregulation.
The purple line represents Downregulation.
The orange line represents Activation.
The yellow line represents Inhibition.
The gray line represents Unclear.
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