JCS02986 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure 1 -

  Fig. S1. Immunocytochemical localization of TRPL and TRPL-eGFP. Cross sections through the eyes of transgenic flies (yw; trpl-eGFP) raised in the dark (left panel) or in the light (right panel) were incubated with rhodamin-conjugated wheat germ agglutinin, which stains the rhabdomeres (red), and with antibodies against TRPL (blue). In addition, TRPL-eGFP fluorescence is shown in green. The overlay of all three channels is depicted in white (upper panel), an overlay of the red and the blue channel appears turquoise (middle panel) and an overlay of the red and the green channel appears yellow (lower panel). Bar, 4 μm. Immunocytochemistry was carried out essentially as described previously (Bähner et al., 2002), except that dissected eyes were fixed for 1 hour instead of 2 hours, 0.3% Triton X-100 was included in the blocking buffer instead of Saponin, and Cy5-coupled secondary antibodies were used.

• Supplemental Figure 2 -

  Fig. S2. Immunocytochemical localization of TRPL and TRPL-eGFP compared with eGFP fluorescence at different times of illumination (A) or dark-adaptation (B). (A,B, upper panels) Cross sections through the eyes of transgenic flies (yw; trpl-eGFP) were incubated with antibodies against TRPL (blue). In addition, TRPL-eGFP fluorescence is shown in green. The overlay of the two channels is depicted in turquoise. Scale bar, 4 μm. (A,B, lower panels) At the indicated time points fluorescence images of intact eyes of flies expressing TRPL-eGFP (yw; trpl-eGFP) were obtained using the water immersion technique. Bar, 15 μm.

• Supplemental Figure 3 -

  Fig. S3. TRPL-eGFP translocation in different light intensities. Transgenic flies (yw; trpl-eGFP) were either dark-adapted or light-adapted with orange light of 450, 10, or 2 Lux, as indicated, for 16 hours. (A-E) Representative images of the eGFP-fluorescence in intact eyes obtained by the water immersion technique. For illumination with 2 Lux images from two individuals are shown that reveal different levels of TRPL-eGFP internalization. (F) The percentage of TRPL-eGFP present in the rhabdomeres was determined. Mean values ± s.d. of at least five experiments are shown.