DOI: 10.6084/m9.figshare.28888943.v1
title: <b>Pyruvate–GPR31 axis </b><b>induces LysoDC dendrite protrusion to M-cell pockets for effective immune responses; Supplementary videos</b>

description: <b>Supplementary Video 1. Two-photon microscopic imaging of a Peyer's patch (PP) explant prepared from untreated </b><b><i>Gpr31b</i></b><sup><strong>+/+</strong></sup><b> </b><b><i>Cx3cr1</i></b><sup><strong>gfp/+</strong></sup><b> mice.</b> PP explants prepared from untreated <i>Gpr31b</i><sup>+/+</sup> <i>Cx3cr1</i><sup>gfp/+</sup> mice were stained with anti-GP2 antibody (red) and vertically monitored with a <i>z</i>-step interval of 0.5 µm by two-photon microscopy. Arrows indicate LysoDC dendrites in M-cell pockets. The arrows remain in place after the image has moved to a depth where the dendrites are not visible. Scale bar, 50 µm. Video corresponds to Figure 2(a).<b>Supplementary Video 2. Two-photon microscopic imaging of a PP explant prepared from untreated </b><b><i>Gpr31b</i></b><sup><strong>-/-</strong></sup><b> </b><b><i>Cx3cr1</i></b><sup><strong>gfp/+</strong></sup><b> mice.</b> PP explants prepared from untreated <i>Gpr31b</i><sup>-/-</sup> <i>Cx3cr1</i><sup>gfp/+</sup> mice were stained with anti-GP2 antibody (red) and vertically monitored with a <i>z</i>-step interval of 0.5 µm by two-photon microscopy. Arrows indicate LysoDC dendrites in M-cell pockets. The arrows remain in place after the image has moved to a depth where the dendrites are not visible. Scale bar, 50 µm. Video corresponds to Figure 2(a).<b>Supplementary Video 3. Two-photon microscopic imaging of a PP explant prepared from untreated </b><b><i>Gpr31b</i></b><sup><strong>+/+</strong></sup><b> </b><b><i>Cx3cr1</i></b><sup><strong>gfp/gfp</strong></sup><b> mice.</b> PP explants prepared from untreated <i>Gpr31b</i><sup>+/+</sup> <i>Cx3cr1</i><sup>gfp/gfp</sup> mice were stained with anti-GP2 antibody (red) and vertically monitored with a <i>z</i>-step interval of 0.5 µm by two-photon microscopy. Arrows indicate LysoDC dendrites in M-cell pockets. The arrows remain in place after the image has moved to a depth where the dendrites are not visible. Scale bar, 50 µm. Video corresponds to Figure 2(a).<b>Supplementary Video 4. Two-photon microscopic imaging of a PP explant prepared from pyruvate-treated </b><b><i>Gpr31b</i></b><sup><strong>+/+</strong></sup><b> </b><b><i>Cx3cr1</i></b><sup><strong>gfp/+</strong></sup><b> mice.</b> PP explants prepared from pyruvate-treated <i>Gpr31b</i><sup>+/+</sup> <i>Cx3cr1</i><sup>gfp/+</sup> mice were stained with anti-GP2 antibody (red) and vertically monitored with a <i>z</i>-step interval of 0.5 µm by two-photon microscopy. Arrows indicate LysoDC dendrites in M-cell pockets. The arrows remain in place after the image has moved to a depth where the dendrites are not visible. Scale bar, 50 µm. Video corresponds to Figure 2(a).<b>Supplementary Video 5. Two-photon microscopic imaging of a PP explant prepared from pyruvate-treated </b><b><i>Gpr31b</i></b><sup><strong>-/-</strong></sup><b> </b><b><i>Cx3cr1</i></b><sup><strong>gfp/+</strong></sup><b> mice.</b> PP explants prepared from pyruvate-treated<i> Gpr31b</i><sup>-/-</sup> <i>Cx3cr1</i><sup>gfp/+</sup> mice were stained with anti-GP2 antibody (red) and vertically monitored with a <i>z</i>-step interval of 0.5 µm by two-photon microscopy. Arrows indicate LysoDC dendrites in M-cell pockets. The arrows remain in place after the image has moved to a depth where the dendrites are not visible. Scale bar, 50 µm. Video corresponds to Figure 2(a).<b>Supplementary Video 6. Finger-like dendrite of LysoDCs in M-cell pockets.</b> PP explants prepared from untreated <i>Gpr31b</i><sup>+/+</sup> <i>Cx3cr1</i><sup>gfp/+</sup> mice were monitored as described in Supplementary video 1. Volume rendering-based 3D reconstruction of the LysoDC protruding a finger-like dendrite into an M-cell pocket is shown. Video corresponds to Figure 2(e).<b>Supplementary Video 7. Spherical dendrite of LysoDCs in M-cell pockets.</b> PP explants prepared from pyruvate-treated <i>Gpr31b</i><sup>+/+</sup> <i>Cx3cr1</i><sup>gfp/+</sup> mice were monitored as described in Supplementary video 4. Volume rendering-based 3D reconstruction of the LysoDC protruding a spherical dendrite into an M-cell pocket is shown. Video corresponds to Cell #2 in Figure 2(g).