Skip to main content
. Author manuscript; available in PMC: 2017 Sep 1.
Published in final edited form as: Cell Signal. 2016 May 17;28(9):1325–1335. doi: 10.1016/j.cellsig.2016.05.013

Fig. 2.

Fig. 2

Spreading and migration in Control and EHD1-KO BMDMS. (A) Microscopy analysis of macrophage cell surface area after 10 minutes CSF-1 stimulation to assess cell spreading. Ehd1fl/fl; CreERT2 BMDMs cultured without (Control) or with (EHD1-KO) TAM were deprived of CSF-1 for 16 hours, stimulated with 100 ng/ml CSF-1 for 10 minutes. Cells were then fixed, permeabilized, stained for polymerized actin using conjugated phalloidin-594, and visualized by confocal microscopy. (B) Quantification of cellular spreading from Fig. 2A was done by analyzing the cell surface area (corresponding to phalloidin staining) using ImageJ software (n = 100 cells were counted in 3 independent experiments. Cell surface area was normalized to Control BMDMs. (C) Trans-well migration assay. Control and EHD1-KO BMDMs were deprived of CSF-1 for 4 hours and allowed to migrate towards CSF-1 (30 ng/ml) in the lower chamber of Boyden Transwell chambers. Filters were HEMA stained and photographed at 20x. (D) Quantification of migration assay from Fig. 2C. Total cells migrated were normalized to Control BMDMs. Data from three independent experiments are presented as the mean ± SEM (*=p<0.05).