Figure 1.

Involvement of STX11 in lipid metabolism

(A) H&E staining and IHC staining of liver sections of C57BL/6J mice fed with a control diet or HFD for 90 days. IHC staining with the anti-STX11 antibody. LDs (black arrows).

(B) Quantitative analysis of STX11 staining and LDs.

(C) Endogenous STX11 protein in the liver from mice treated with a control diet or HFD for 90 days was measured by immunoblotting. Perilipin2, a lipid droplet-specific protein marker. β-tubulin was used as the loading control.

(D) Cellular distribution of STX11 protein assessed by immunoblotting analysis. 1, HepG2; 2, THLE-2; 3, HeLa; 4, HK2; 5, HEK293T. β-tubulin was used as the loading control.

(E) Relative expression of STX11 in different cells.

(F and G) Overexpression of STX11 affects LD accumulation in cells. STX11-Myc proteins immunofluorescence staining with anti-Myc (red) antibodies was performed to reveal LD changes in THLE-2 and HeLa cells overexpressing STX11-Myc. Cells were incubated under normal growth conditions with 400 μM OA complexed to albumin for 24 h. Lipid droplets were stained with BODIPY 493/503 fluorescent dye. Quantification of LDs under the fluorescence microscope (G) (n = 6 cells for Ctrl; n = 6 cells for STX11 overexpression).

(H and I) Knock down of STX11 or simultaneous overexpression of STX11-Myc in THLE-2 and HeLa cells treated as described in (F and G). STX11 proteins measured by immunoblotting using STX11 antibodies (I). β-actin was used as the loading control.

(J) Relative intensity of LDs under the fluorescence microscope. Data are shown as mean ± SEM (n = 3). ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, t test.