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. 2019 Jan 21;5(4):559–569. doi: 10.1021/acsinfecdis.8b00301

Figure 5.

Figure 5

ARL8B is a bonafide LD associated protein. (a) ARL8B-mCherry colocalization with LAMP1+ organelles (lysosomes) in RAW264.7 cells. Scale bar = 10 μm. (b) Quantification of colocalization rate of ARL8B-mCherry and LAMP1 in RAW264.7 cells. (c) mCherry and ARL8B-mCherry (red) localization relative to lipid droplets stained with BODIPY665/676 (pseudocolored in green) in RAW264.7 cells loaded with oleic acid. The left panel shows the maximum intensity projection while the middle and right panel show indicated z stacks of a SIM image. Scale bar = 5 μm. (d) Thin layer chromatogram of lipids isolated from indicated fractions: total cell lysate (T) and LD fraction (LD). 0.2% of total cell lysate while 4% of the LD fraction was loaded. Standards for diacylglycerol (DAG), cholesterol (C), cholesterol ester (CE), and triacylglycerol (TG) are indicated. (e) Immunoblotting for mCherry, LAMP1, and PLIN2 in LDs isolated from RAW264.7 cells in (b) and (c). * represents possible degradation product of ARL8B-mCherry.