TMT analysis of LARP6 knockdown impact on protrusion induced changes in the proteome of MDA-MB231 cells

To assess if LARP6 is required for upregulation of ribosomal proteins upon protrusion induction, MDA-MB231 breast cancer cells were treated with non-targeting (NT) or LARP6 siRNA for 48 hrs, before being seeded on top of collagen-I coated 3 micron transwell filters (corning). No media was added to the bottom chamber and cells were allowed to adhere to filter overnight. The next day, the media on top of cells was refreshed, and the filters were either left with no media in the bottom chamber (closed pores), or with media also added to the bottom chamber (open pores) in order to allow formation of protrusions. Cells were allowed to form protrusions for 12 hrs before being lysed and analysed by mass spectrometry.