Figure 7.

AP006216.5, a human ortholog of GM47544, promoted the degradation of ApoC3 and thus exerted a lipid-lowering effect.

(A) Schematic representation of the human APOA1/C3/A4/A5 locus.

(B) Relative expression levels of ENST00000439104.1, AP006216.5, and ENST000000457746.1 under PA stimulations in THLE-3 cells were detected by RT-qPCR analysis.

(C) Relative expression of APOA1/C3/A4/A5 in control and AP006216.5-overexpressed THLE-3 cells was measured by RT-qPCR.

(D) APOA1/C3/A4/A5 protein levels in control and AP006216.5-overexpressed THLE-3 cells were measured by western blot.

(E) Oil Red O staining of control and AP006216.5-overexpressed THLE-3 cells.

(F) Intracellular TG and TC (F) levels of control and AP006216.5-overexpressed THLE-3 cells with PA treatment (250 μM, 24 h).

(G) Dil-LDL uptake was quantified under fluorescence microscopy in control and AP006216.5-overexpressed THLE-3 cells treated with Dil-LDL (15 μg/mL) for 24 h. Quantification of the relative fluorescence intensity of internalized DiI-LDL in control and C cells. Red: Dil-LDL; blue: DAPI staining. Scale bar, 100 μm.

(H) Cellular HL activity in control and AP006216.5-overexpressed THLE-3 cells was detected.

(I) THLE-3 was transfected with vectors expressing the FLAG-APOC3, with or without vectors expressing the AP006216.5. IP assays were performed using anti-FLAG antibodies, followed by western blotting analysis with antibodies against ubiquitin.

Data are presented as mean ± SD in B, C, E, F, G, and H, with statistical significance determined with unpaired two-tailed Student's t-test.