Fig. 3.

Accelerated cell spreading in Atgl−/− macrophages. a, d Macrophages from Wt and Atgl−/− mice were plated on fibronectin-coated glass coverslips for 10 min (a) in the absence or (d) presence of SDF-1 (60 ng/ml) and LPA (9 ng/ml) for 2 and 5 min. Adherent cells were fixed and stained with phalloidin AlexaFluor-586. Images were taken using a Leica AP5 AOBS confocal microscope. Arrow in (a) points to broad protrusions in an Atgl−/− macrophage. Scale bars 5 μm. b, e Cell spreading was calculated by counting the percentage of spread cells. c, f Mean cellular area was measured using MetaMorph software. Data from two independent experiments are presented as mean values of >650 cells per genotype ±SEM. g Fluorescence intensity of phalloidin AlexaFluor-586-stained Wt and Atgl−/− macrophages was measured by flow cytometry. The histograms show untreated cells (brown) and macrophages treated with LPA for 2 min (green), 5 min (yellow) and 10 min (blue). *p < 0.05, **p ≤ 0.01, ***p ≤ 0.001