Figure 3.

STX11 directly interacts with ATGL

(A) Schematic representation of the structure of murine ATGL, its mutants, and the PT domain in ATGL is indicated as the gray region.

(B) STX11 interacts with the PT domain of ATGL. HEK293T cells were co-transfected with STX11-HA together with different ATGL constructs (ATGL-Myc + STX11-HA, ATGL-PT-Myc + STX11-HA, ATGL-ΔPT-Myc + STX11-HA). STX11-HA proteins were immunoprecipitated with anti-HA antibodies. STX11 and ATGL proteins in immunoprecipitated lysates were detected by immunoblotting with HA and Myc antibodies.

(C) Schematic representation of the structure of murine STX11, its mutants, and the Habc domain and the SNARE motif in STX11 are indicated as the gray region.

(D) C-terminal of STX11 interacts with the PT domain of ATGL. HEK293T cells were co-transfected with STX11ΔC-HA and ATGL-PT-Myc. Other groups and immunoprecipitation and immunoblotting analyses were performed as described in (B).

(E) STX11 co-localizes with PT domain of ATGL. Immunofluorescence staining of STX11-HA and ATGL-PT-Myc in HeLa cells was performed using anti-HA (green) and anti-Myc (red) antibodies. Intensity traces from the original images were plotted using ImageJ software. Curve coincidence illustrates that the two proteins have co-localized in theory.

(F) HeLa cells were co-transfected with STX11ΔC-HA and ATGL-Myc. STX11ΔC-HA proteins immunofluorescence staining was performed by using anti-HA (green) and ATGL-Myc proteins were stained with anti-Myc (red) antibodies.

(G and H) GST pulldown analysis between recombinant STX11-GST, ATGL-MBP-His, and ATGL-MBP-PT-His.