Pyruvate–GPR31 axis induces LysoDC dendrite protrusion to M-cell pockets for effective immune responses; Supplementary videos

Supplementary Video 1. Two-photon microscopic imaging of a Peyer's patch (PP) explant prepared from untreated Gpr31b+/+ Cx3cr1gfp/+ mice. PP explants prepared from untreated Gpr31b+/+ Cx3cr1gfp/+ mice were stained with anti-GP2 antibody (red) and vertically monitored with a z-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).Supplementary Video 2. Two-photon microscopic imaging of a PP explant prepared from untreated Gpr31b-/- Cx3cr1gfp/+ mice. PP explants prepared from untreated Gpr31b-/- Cx3cr1gfp/+ mice were stained with anti-GP2 antibody (red) and vertically monitored with a z-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).Supplementary Video 3. Two-photon microscopic imaging of a PP explant prepared from untreated Gpr31b+/+ Cx3cr1gfp/gfp mice. PP explants prepared from untreated Gpr31b+/+ Cx3cr1gfp/gfp mice were stained with anti-GP2 antibody (red) and vertically monitored with a z-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).Supplementary Video 4. Two-photon microscopic imaging of a PP explant prepared from pyruvate-treated Gpr31b+/+ Cx3cr1gfp/+ mice. PP explants prepared from pyruvate-treated Gpr31b+/+ Cx3cr1gfp/+ mice were stained with anti-GP2 antibody (red) and vertically monitored with a z-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).Supplementary Video 5. Two-photon microscopic imaging of a PP explant prepared from pyruvate-treated Gpr31b-/- Cx3cr1gfp/+ mice. PP explants prepared from pyruvate-treated Gpr31b-/- Cx3cr1gfp/+ mice were stained with anti-GP2 antibody (red) and vertically monitored with a z-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).Supplementary Video 6. Finger-like dendrite of LysoDCs in M-cell pockets. PP explants prepared from untreated Gpr31b+/+ Cx3cr1gfp/+ 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).Supplementary Video 7. Spherical dendrite of LysoDCs in M-cell pockets. PP explants prepared from pyruvate-treated Gpr31b+/+ Cx3cr1gfp/+ 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).