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Additional Supporting Information may be found in the online version of this article.

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CNE_22773_sm_SuppFig2.tif3264KSupplementary Figure 2. (A magenta green version of Figure 3 from the manuscript for the assistance of color-blind readers.) IRBP distribution in saline washed chicken retina. Retinas were washed three times, flatmounted, and probed with mAb F7 anti-IRBP followed by goat anti-mouse IgG-647. Overview of merged confocal fluorescence images at 633 nm (red, IRBP), and 488 nm (green, oil droplet auto-fluorescence). (A) En face orientation of cone array of IRBP indirect immunofluorescence (arrow, double-cone). (B) En face orientation incubated without primary antibody. (C) Oblique section showing inner segment (IS) and outer segment (OS) zones (double arrows). Horizontal arrows, distal cone outer segments tips. (D) Enlargement of marked double-cone region in panel A. Arrowheads, ring-like association of IRBP to cone outer segments; asterisk, region between IRBP cone-associated domains; scale bar in panel D is 3.3 μm, and represents 10 μm for panels A-C.
CNE_22773_sm_SuppFig4.tif3373KSupplementary Figure 4. (A magenta green version of Figure 5 from the manuscript for the assistance of color-blind readers.) Localization of IRBP and PNA-binding glycans in the pig retina. Isolated retinas were probed with: (A) mAb F7 anti-IRBP followed by goat anti-mouse IgG-647; (B) PNA-488. Merged fluorescence is shown in panel C. IRBP and PNA-binding glycans show strong co-localization (arrow). Asterisk, cone matrix profile; Scale bar in panel C is 10 μm.
CNE_22773_sm_SuppFig5.tif1828KSupplementary Figure 5. (A magenta green version of Figure 6 from the manuscript for the assistance of color-blind readers.) Isolated turkey cones probed with PNA-488 (A-C), or WGA-647 (D-F). Washed retinas were incubated with either lectin prior to gentle trituration to generate CIS/COS. (A,D) Retained cone matrix sheath (arrowheads) is detectable around the outer and inner segments by DIC. (B,E) Confocal immunofluorescence of PNA-488 and WGA-647 fluorescence, respectively. (C,F) Merged fluorescence and DIC. OS, outer segment; OD, oil droplets; Ellip, Ellipsoid; Myd, myoid; arrow, PNA-positive cone matrix sheath. Scale bar in panel F is 5 μm.
CNE_22773_sm_SuppFig6.tif1950KSupplementary Figure 6. (A magenta green version of Figure 7 from the manuscript for the assistance of color-blind readers.) Localization of IRBP and PNA-binding glycans in dissociated turkey photoreceptors. Washed retinas were incubated with mAb F7 anti-IRBP followed by goat anti-mouse IgG-647. The tissue was then probed with PNA-488, and gently triturated to generate photoreceptor clusters or CIS/COS. Panels A-C show a single cone (outer segment marked with arrow), and nearby rod outer segment (arrowhead); Panels D-F show a double cone. (A,D) Cone matrix sheath labeling with PNA-488; (B,E) IRBP localization within the cone matrix sheath; (C,F) Merged corresponding fluorescence and DIC images. Note that IRBP co-localization within the cone matrix sheath is restricted to an outer segment subdomain (yellow merged fluorescence). Ellip, Ellipsoid; OD, oil droplet; scale bar in panel F represents 5 μm for panels A-C, and 8.7 μm for panels D-F.
CNE_22773_sm_SuppFig7.tif6265KSupplementary Figure 7. (A magenta green version of Figure 8 from the manuscript for the assistance of color-blind readers.) Localization of IRBP and PNA-binding glycans in isolated chicken IPM. Matrix sheets were delaminated from retinas in 2 mM CaCl2 and mounted onto subbed slides. (A) Indirect IRBP immunofluorescence using mAb F7 anti-IRBP detected with goat anti-mouse IgG-647; (B) Distribution of PNA-488 binding glycans; (C) Merged fluorescence. Asterisk, matrix appearing to bridge between cone matrix sheath structures; Arrows, fluorescence co-localization in cone matrix sheaths. Scale bar in panel C is 10 μm.
CNE_22773_sm_SuppFig9.tif1786KSupplementary Figure 8. (A magenta green version of Figure 9 from the manuscript for the assistance of color-blind readers.) Localization of IRBP, and PNA- and WGA-binding glycans within isolated turkey IPM. The matrix sheet was delaminated from a freshly isolated retina in 2 mM CaCl2 and probed with: (A) MAb F7 anti-IRBP followed by goat-anti mouse IgG-647; (B) PNA-488; and (C) WGA-555. (D) Merged IRBP, PNA, and WGA fluorescence. IRBP immunofluorescence is shown in blue because three fluorophores are used (647-blue, 633nm; 488-green, 488nm; 555-red, 561nm). Asterisk, WGA positive matrix appearing to bridge nearby cone sheaths. Arrows, co-localization of IRBP and PNA/WGA-binding glycans. Arrowheads, rod-associated matrix. Scale bar in panel D is 5 μm.

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