Fig. 3.

ECF21 family sigma factor, Das1, is required for hypervesiculation. (A) Coomassie blue stain comparing protein profiles between WT, Δdas1, Δdma1, and Δdas1-dma1. This shows that deletion of Das1 in the Δdma1 background restores WT levels of vesiculation. Samples were normalized by OD600 values and run on 10% SDS-PAGE gel. (B) Schematic of constructs used for targeted pulldown assay (created with https://BioRender.com). (C) Western blots using anti-polyHis (Green) and anti-GST (Red). Top: Gel shows that equivalent amounts of bait and prey proteins were mixed prior to passage through a column containing anti-GST resin. Bottom: Gel showing the proteins present after the elution step, which demonstrates that the N terminus of Dma1 is sufficient to sequester Das1.