Deciphering the role of SRSF5 in enteroviral replication

Abstract

We previously conducted RNA-interactome and identified several members of the SR protein family. Knockdown experiments revealed that several SR proteins are involved in the EV-A71 replication, including SRSF5. This study looks at how SRSF5 is expressed using Western blot assays for plasmid transfection and siRNA-mediated knockdown, rescue, and overexpression. In the plasmid assay, faint signals that correspond to the SRSF5 proteins encoded by the mCherry-tagged plasmid appeared at the expected size (75-63 kDa), showing problems with transfection. Smudged bands and unclear alpha-tubulin signals pointed to errors in sample preparation and gel handling. The complete assay showed successful SRSF5 overexpression, proving the plasmid works. However, knockdown results showed SRSF5 was not successfully knocked down, which might be due to the poor transfection this time. Future improvements should focus on better transfection methods, protein preparation, and siRNA design. Despite this limitation, these works contribute to assessing the role of SRSF5’s in EV-A71 infection.