Identification of GM47544 as a hepatic regulator of lipid metabolism.
(A) Representative heatmap showing the response of 12 differentially expressed lncRNAs in mice liver to HFD-feeding. Four highly expressed mRNAs (fold change > 2; FDR-adjusted P < 0.05) located in the ApoA1/C3/A4/A5 gene cluster in the livers of HFD mice. The heatmap was drawn based on normalized expression levels (n = 5).
(B) Schematic representation of the mouse ApoA1/C3/A4/A5 gene cluster locus.
(C) Relative RNA expression of GM47544 in the livers of wide type (WT) or db/db male mice (n = 8).
(D) Relative RNA expression of GM47544 in AML-12 cells with the treatment of PA (250 μM) for 24 h.
(E) Relative RNA expression levels of GM47544 in multiple mouse tissues (n = 6).
(F) RNA FISH analysis of GM47544 in AML-12 cells revealed that the majority of GM47544 is located in the cytoplasm. Red: GM47544; blue: DAPI staining. Scale bar, 0.5 μm.
(G) Detection of GM47544 in the cytoplasm and nucleus of AML-12 cells by qPCR. As a control, GAPDH was used for the cytoplasm and U6 was used for the nucleolus.
(H) Relative expression levels of GM47544 in AML-12 cell lines stably overexpressed GM47544 (OE-GM47544) and the negative control (OE-Ctrl).
(I) Gene set enriched expression (GSEA) plots for fatty acid metabolic pathways. GSEA was performed based on average GM47544 expression (OE-GM47544 versus OE-Ctrl).
(J) Oil Red O staining and hematoxylin staining of lipid droplets in control and GM47544-overexpressed AML-12 cells treated with PA (250 μM, 24 h). Scale bar, 100 μm.
(K) Intracellular TG and TC levels of control and GM47544-overexpressed AML-12 cells with or without PA treatment (250 μM, 24 h).
(L) Left: Dil-LDL uptake was quantified under fluorescence microscopy in control and GM47544-overexpressed AML-12 cells treated with Dil-LDL (15 μg/mL) for 24 h. Red: Dil-LDL; blue: DAPI staining. Scale bar, 100 μm. Right: quantification of the relative fluorescence intensity of internalized DiI-LDL in control and GM47544-overexpressed AML-12 cells. (M) Cellular HL activity in control and GM47544-overexpressed AML-12 cells was detected.
Data are presented as mean ± SD in C, D, H, J, K, L, and M, with statistical significance determined with unpaired two-tailed Student's t-test. Data are presented as mean ± SD in E and G.