Fig 1: CDK5 and 14-3-3ε and KIAA0528 are found on lipid droplet (LD) cargo. Intact purified LDs were probed with antibodies against CDK5, 14-3-3ε, KIAA0528, and GFP (negative control), the percentage of LDs that had detectable immunofluorescence signal quantification is shown in a and sample immunofluorescence images are shown in b with 2 μm scale bars, and white arrows designate lipid droplets. To assess CDK5 localization to LDs, proteins were precipitated from purified LDs and analyzed by WB. Perilipin-2: (an LD marker) is present on both samples and 25× CDK5 is present on the LDs from the CDK5 overexpression background (c). To assess 14-3-3ε and KIAA0528 localization, IF experiments were performed on LDs purified from wild-type or siRNA-treated cells. LDs from the KD cells showed reduced levels of the respective proteins, quantified in d, with example images for each experiment in the supplement, Fig. 6. To further confirm KIAA0528 LD localization, LDs were purified from wild-type (WT) or KIAA0528-GFP overexpressing cells, and the percentage of LDs with GFP puncta was quantified. LDs from KIAA0528-GFP cells show increased puncta compared to WT cells (e). Representative fluorescence images are in the supplement, Fig. 6. Quantified data represent the mean ± s.e.m. of n = 2 independent experiments. Two-sided test of proportions P values *P < 0.05, ***P < 0.001, ****P < 0.0001