Fig. 1: ApoL6 is a LD-associated protein and enhances triglyceride accumulation.
From: ApoL6 associates with lipid droplets and disrupts Perilipin1-HSL interaction to inhibit lipolysis
a RT-qPCR and Northern blotting for ApoL6 mRNA levels in various tissues. (n = 3 mice, **p = 0.0061, **p = 0.0019, ***p < 0,001, two-tailed Student’s t test compared to expression levels in liver). Data are expressed as mean ± SD. b RT-PCR and RT-qPCR for ApoL6 mRNA levels in stromal vascular and adipocyte fractions of WAT (n = 4 mice, ***p < 0.001, two-tailed Student’s t test). Data are expressed as mean ± SD. c RT-qPCR (left) and immunoblotting (right) during 3T3-L1 adipocyte differentiation. d Northern blotting (left) and immunoblotting (right) of WAT. e Northern blotting of WAT. f Immunofluorescence of differentiated 3T3-L1 cells infected with ApoL6-GFP (green) adenovirus stained with (red), (bar = 10 μm) and immunoblotting after sucrose separation (right). g Immunoblotting after incubation of lipid membrane strips with either WT ApoL6-GST or control GST (right and middle) or mutated ApoL6 -GST (Mu-ApoL6-GST, aa286-292, from RYRKLQR to DYDDLQD, right). h Immunoblotting after infection of adenoviral ApoL6 in differentiated 3T3-L1 adipocytes, image (bar=10 μm), quantification of LD size and TAG levels (right, n = 5, ***p < 0.001, two-tailed Student’s t test). Data are expressed as mean ± SD. Experiment was repeated twice. i Human adipocytes differentiated from human fibroblasts were infected with lentiviral h-ApoL6-HA. Immunoblotting with h-ApoL6 antibody, cell images (bar = 10 μm), and TAG levels (n = 6, ***p < 0.001, two-tailed Student’s t test). j Immunoblotting after scramble and ApoL6-shRNA infection in 3T3-L1 adipocytes. LipidTOX staining (bar = 10 μm), quantification of LD sizes and TAG levels (n = 6. **p = 0.0025, two-tailed Student’s t test). Data represent mean ± SD. Independent experiments were repeated twice. k Human adipocytes were infected with lentiviral ApoL6 shRNA. Immunoblotting after infection, cell images (bar = 10 μm) and TAG levels (n = 6, **p = 0.003, two-tailed Student’s t test). l Lipids were extracted from [U-14C-TAG] pulse-labeled 3T3-L1 adipocytes and separated by TLC. Image for TLC plate. 14C-TAG was quantified by scintillation counting (n = 4, ***p < 0.001, two-tailed Student’s t test). DAG levels were measured in lipids extracted from 3T3-L1 adipocytes (n = 6, ns p = 0.0912, two-tailed Student’s t test) and WAT from mice (right, n = 4, ns p = 0.058, two-tailed Student’s t test). Data represent mean ± SD. Source data are provided as a Source Data file.
