Figure 4.

RalB lysine fatty acylation promotes A549 cell migration. (A) Images of the stained Boyden chambers after 24 hour cell migration. For the experiment cells were starved of serum for 12hr, prior to being counted and seeded in Boyden chambers. Experiments were done in at least biological duplicates and images shown are from one representative experiment. Images were taken on a Motic AE21 microscope, using a 20X objective. Motic Images Plus 2.0 software was used to obtain the images. (B) Quantification for experiments shown in A. Quantification was done by measuring the absorbance of the crystal violet stain of migrated cells and normalizing to the starting cell number as determined by cell titer blue. (C) Western blots to confirm SIRT2 KD, and Flag-RalB WT and Flag-RalB 8KR overexpression in A549 cells.