Figure 3.

Absence of GRAB does not affect Rab8a recruitment or downstream TLR signalling. (a) Live cell confocal spinning disc imaging of LPS-treated control and GRAB KO cells transiently overexpressing GFP-Rab8a showing recruitment to macropinosomes and tubules. Quantification of Rab8a retention on macropinosomes was measured by the total number of timeframes Rab8a is spent enriched on each macropinosome, and 5 cells of each cell line was used for quantification (n = 5). Scale bars, 10 µm. (b) Immunoblotting for levels of Akt phosphorylation using phospho specific antibodies on the lysates of control and GRAB KO cells treated with LPS over a 60 min time course. The immunoblot is representative and levels of Akt phosphorylation was quantified using the densitometry ratio between the band intensities of GAPDH and phospho-Akt levels. Significance was measured via two-way analysis of variance (ANOVA) (n = 3)