Fig. 1: Confocal microscopy and CLEM revealed the ring-like structure of the Httex1 72Q inclusions in HEK cells. | Nature Communications

Fig. 1: Confocal microscopy and CLEM revealed the ring-like structure of the Httex1 72Q inclusions in HEK cells.

From: Nuclear and cytoplasmic huntingtin inclusions exhibit distinct biochemical composition, interactome and ultrastructural properties

Fig. 1

a Epitope mapping of the Httex1 antibodies used in this study. b Confocal imaging of Httex1 72Q inclusions formed 48 h after transfection in HEK cells. All the Htt antibodies showed strong immunoreactivity to the periphery of the inclusions. The nucleus was counterstained with DAPI and the F-actin with phalloidin. White arrows indicate the colocalization of the F-actin with the ring-like structure of Httex1 inclusions. Scale bar = 20 μm and 10 μm. c, d CLEM of Httex1 72Q inclusions. c Confocal imaging of Httex1 72Q inclusions formed 48 h after transfection in HEK cells. The selected area (white square) was examined by EM (d). Orthogonal projection (Orth. Pr.), Scale bars = 5 μm. d The presence of membrane fragments and vesicles are indicated by the white arrowheads. The binary image (inset) shows the core and the shell ultrastructure of the Httex1 72Q inclusion. Scale bar = 500 nm. e Representative EM images of Httex1 72Q inclusion. Higher magnification (white square) are shown in the right panel. Dashed lines delimit the aggregate and the core of the inclusion. Internalized membranous structures and the mitochondria are indicated by the orange and the green arrowheads, respectively. Scale bar = 1 μm and 500 nm. f 3D model of Httex1 72Q cellular inclusion and surrounding organelles (top view). The Httex1 inclusion body (IB) shell is represented in purple, the core in cyan. ER membranes are shown in green, intra-inclusion membranous structures in white, the nucleus in blue, mitochondria in yellow, and the stacked ER cisternae are indicated by a red arrowhead. Scale bar = 2 μm. g Representative EM images of Httex1 39Q inclusion formed 48 h after transfection in HEK cells. The white square indicates the area shown in the right panel at higher magnification. Dashed lines delimit the inclusion. Scale bar = 1 μm and 500 nm. h 3D model of the Httex1 39Q IB is shown in purple, surrounded by mitochondria (yellow), ER structures (green), stacked ER cisternae (red arrowheads), intra-inclusion membranous structures are shown in white and the nucleus in blue. Scale bar = 2 μm.

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