Antibody Staining Using antibody staining, you observe that your protein of inte

Question

Antibody Staining

Using antibody staining, you observe that your protein of interest localizes as shown in the picture to the right (the white regions).

1. What can you conclude about localization of the mRNA at this stage? (1 pt)

2. Give TWO reasons for this conclusion. (1 pt)

Antibody Staining Using antibody staining, you observe that your protein of interest localizes as shown in the picture to the right (the white regions). 1. What can you conclude about localization of the mRNA at this stage? (1 pt) 2. Give TWO reasons for this conclusion. (1 pt)

Explanation / Answer

1. Proteins are known to function in microRNA (miRNA)-based translational repression, we lack a comprehensive understanding of temporal relationships between the mRNA, miRNA and their constituent proteins. To understand the dynamics of miRNA and protein interactions, we created a synthetic inducible miRNA system in mammalian cells. By visualizing single mRNAs and observing their co-localization with proteins over time, we produced a temporal association map of miRNA-associated factors. Argonaute2, Dcp1a, hedls and Rck co-localize with miRNA-regulated mRNA after 24 h of miRNA induction, and RNAi knockdown of any one of these proteins affected the co-localization of any of the other proteins with miRNA-regulated mRNA, demonstrating that these proteins could interact with each other in a complex. We identified Argonaute2 and hedls as proteins that co-localize and interact with miRNA-regulated mRNA, indicating that processing body components are involved in long-term storage of miRNA-regulated mRNA.

2. Protein is localized and antibody is appeared

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