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. Author manuscript; available in PMC: 2014 Sep 1.
Published in final edited form as: Mol Cell Neurosci. 2013 Jul 10;0:283–297. doi: 10.1016/j.mcn.2013.07.002

Figure 1. BV2 cells proliferation and morphology are altered with or without FBS.

Figure 1

A. Phalloidin (PHL-green) and GSA-lectin (red) labeling of cells grown in Hyclone; ADCF-mAb (HyC-serum free) media and cells grown in 10% FBS (D10%). B. Quantification of the IF shows 85% of HyC cells are processed while 90% of D10% cells are round. C. MTT proliferation assay shows significantly increased rate of proliferation of D10% cells compared to HyC 4 days after seeding. Assay was performed a minimum of 3 times. D–E. HyC or D10% cells labeled with cholera toxin (Ctx-green) to stain PM lipid rafts. HyC cells are elongated and processed (left panels) while D10% cells are rounded (i, ii). Ctx specificity for PM was tested with the cytoplasmic microglia marker, Iba1 (E). Magnification bars = 10μM. Statistical analysis was performed with student’s paired t-test. F. TEM analysis of BV2 cells. Small A (a) and C are D10% grown cells. B (b) and D are HyC grown cells. A. Numerous mitochondria (M) and a dense network of rough endoplasmic reticulum (RER) is evident, typical of highly proliferative cells. Inset shows a representative caveolae at the PM from these cells. B. HyC grown cells have minimal mitochondria, and reduced RER. Large cytoplasmic vacuoles (V), some with vesicles inside or entering (arrows) are present indicating these cells are still able to internalize and process extracellular proteins. Inset shows typical HyC caveolae. (Arrow) marks a potential vesicle fusion with a cytoplasmic vacuole. C and D show representative vesicle uptake in either culture condition. Both clathrin coated vesicles (CCV) and morphological caveolae are seen in both images (arrowheads = CCV; arrows = caveolae). One degradative multi-vesicular body (MVB) is seen in D. * magnification bar = 200nm ** magnification bar = 50nm. G. O2 consumption rate (OCAR) is enhanced in BV2 cells grown with D10% media suggesting higher mitochondrial respiratory capacity than HyC cells. Glycolysis, measured by extracellular acidification rate (ECAR), is also higher at baseline in D10% cells compared to HyC.