Fig. 6: Depletion of Tctn1 causes increased diffusion rates at the transition zone.

a Airyscan images from isolated iCre/Tctn1^het and iCre/Tctn1^flox outer segments stained with anti-Bbs5 or anti-IFT20 antibodies (magenta) along with α-tubulin (green) and WGA (gray). Averaged intensity plots are shown to the right. The intensity of either Bbs5 or IFT20 (10–15 transition zones analyzed per biological replicate, n = 3). Error Bars indicate S.D. Statistical significance was determined with an unpaired two-tailed t test. BBS5 ns, p = 0.3428 and IFT20 ns, p = 0.2749. b Representative images from wild type mouse retinas co-electroporated with Rho-GFP (green) and mKate-Cetn2 (magenta). Low magnification to the left (Scale bar, 5 µm); high magnification and contrast images to the right (Scale bars, 2 µm). Magenta box show representative ROI used for photobleaching the transition zone of transfected rods. Nuclei are counterstained with DAPI (blue). c Representative FRAP traces from Tctn1^flox and iCre/Tctn1^flox rods plotted for the change in Rho-GFP fluorescence intensity normalized to a pre-bleached average intensity (∆F/F0) over time in seconds (s). FRAP traces are fitted using a phase I non-linear regression function. d Bar graph plots the average half-time of recovery (T_1/2, s) calculated from FRAP traces from Tctn1^flox (16 cells, n = 5 biological replicates) and iCre/Tctn1^flox (16 cells, n = 7 biological replicates) at 1 month. Error bars indicate S.D. Statistical significance was determined with Welch’s two-tailed t test. *, p < 0.018. e Representative image from wild type mouse retinas electroporated with GFP-TM (green). Low magnification to the left (Scale bar, 5 µm); high magnification and contrast images to the right (Scale bar, 2 µm). Magenta box show representative ROI used for photobleaching the plasma membrane of transfected rods. f Representative GFP-TM FRAP traces from Tctn1^flox and iCre/Tctn1^flox rods. g Bar graph plots the average T_1/2 (s) calculated from FRAP traces from Tctn1^flox (23 cells, n = 4 biological replicates) and iCre/Tctn1^flox (13 cells, n = 4 biological replicates) at 1 month. Error bars indicate S.D. Statistical significance was determined with Welch’s two-tailed t test. ns, p = 0.6547. Source data are provided as a Source Data file.