Figure 8.

Rabin8 and GRAB double KO reduces Akt enrichment on LPS induced Macropinosomes. Control and double KO cells were transiently co-transfected with C1-GFP and TagRFP-T-Akt1 and live cell spinning disc confocal microscopy was performed. A ratiometric ‘heat map’ of Akt intensity was generated by using the ratio of TagRFP-Akt1 signal intensity to the soluble GFP to identify regions of Akt membrane enrichment. The cells were imaged with and without LPS. Macropinosomes with Akt enrichment were counted as number of events per cell, expanded panels show examples of macropinosomes observed with and without Akt enrichment in the control and double KO cells respectively. Movies were taken over 15 min with 5 sec intervals and quantification was performed by counting the number of Akt enriched macropinosomes observed per cell over a sample of 5 cells for each cell line. Significance was measured via Student’s t-test. (*P < 0.05, **P < 0.01 and n = 5 cells). Scale bars, 10 µm