I have examined the effect of a frameshift mutation in different cells derived from patient, such as fibroblast, EBV-Lymphoblastiod cells from the PBMC, Retinal pegmented epithelial cells and PBMC. My results showed variable but significant down regulation of mRNA in all cells except PBMC, the level of mRNA was unchanged. Unfortunately, I don't have enough cell to check for protein level in these cells. A truncated product was observed in the other cell that showed reduction in mRNA. NMD was investigated by cyclohexamide in fibroblasts and RPE which showed resistance of NMD in fibroblast but not in RPE. My question is why we did not see reduction of mRNA in PBMC while EBV-LYB and other cells showed downregulation of the mRNA??
Dear Mohammad, mRNA synthesis is more active in cycling cells. So, you can suppress it in immortalized cells. Against, PBMC are mostly resting cells. For this reason, you can not decrease mRNA synthesis in PBMC so much. I think, after stimulation of PBMC by any mitogen in vitro, you will see much more prominent decrease.
The cells you observed are not representative of the tisuues you tested. Avbsence of what you are looking for does not mean it is does not occur naturally.
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