XB-IMG-136658
Xenbase Image ID: 136658
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Figure 4. GFP-IFT43 but not GFP-IFT20 requires Rsg1 function for localization to peri-basal body IFT pools. (a) A single confocal slice of the apical surface of a control MCC expressing GFP-IFT20 and centrin-RFP. (b) A single confocal slice of a representative Rsg1 KD MCC co-expressing GFP-IFT20 and centrin-RFP. Note that despite the decreased density of centrin-RFP foci there is still a strong correlation between the centrin-RFP and GFP-IFT20 localization patterns. (c) A single confocal slice of a representative Rsg1 KD2 MCC co-expressing GFP-IFT20 and centrin-RFP. (d) A representative control MCC expressing GFP-IFT43 and centrin-RFP. (e) A representative Rsg1 KD MCC expressing GFP-IFT43 and centrin-RFP. Notice the impaired localization of GFP-IFT43 to centrin-RFP foci. (f) A representative Rsg1 KD2 MCC expressing GFP-IFT43 and centrin-RFP. (g) Quantification of the mean of GFP-IFT20 foci mean intensities, as normalized to the same value for centrin-RFP, shows no significant change between control and Rsg1 KD MCCs (Ctl: 0.84 ? 0.28, n = 45 cells, 8 embryos vs. Rsg1 KD: 0.71 ? 0.26, n = 43 cells, 8 embryos, P = 0.042). (h). Quantification of the mean of GFP-IFT43 foci mean intensities, as normalized to the same value for centrin-RFP, shows a significant decrease between control and Rsg1 KD MCCs (Ctl: 0.59 ? 0.14, n = 41 cells, 8 embryos vs. Rsg1 KD: 0.37 ? 0.25, n = 41 cells, 8 embryos. ***P <0.0001). (i) Quantification of the mean area of GFP-IFT20 foci in a cell normalized against the same value for centrin-RFP shows no significant change between control and Rsg1 KD conditions (Ctl: 2.14 ? 0.79, n = 44 cells, 8 embryos vs. Rsg1 KD: 2.41 ? 0.80, n = 41 cells, 8 embryos; P = 0.3477). (j) Quantification of the mean area of GFP-IFT43 foci in a cell normalized against the same value for centrin-RFP shows a significant decrease in Rsg1 KD MCCs as compared to controls (Ctl: 1.98 ? 0.74, n = 39 cells, 8 embryos vs. Rsg1 KD: 0.69 ? 0.46, n = 39 cells, 8 embryos; ***P <0.0001). (k) There is no significant change in the number of GFP-IFT20 foci detected per centrin-RFP foci between control and Rsg1 KD MCCs (Ctl: 0.81 ? 0.15, n = 45 cells, 8 embryos vs. Rsg1 KD: 0.78 ? 0.19, n = 43 cells, 8 embryos; P = 0.062). (l) There is a significant reduction in the number of GFP-IFT43 foci detected per centrin-RFP foci between control and Rsg1 KD MCCs (Ctl: 0.76 ? 0.17, n = 41 cells, 8 embryos vs. Rsg1 KD: 0.25 ? 0.25, n = 41 cells, 8 embryos; ***P <0.0001). (m) There is a reduction in the number of centrin-RFP foci detected on average in Rsg1 KD MCCs as compared to controls (Ctl: 106.90 ? 21.39, n = 45 cells, 8 embryos vs. Rsg1 KD: 72.95 ? 28.63, n = 43 cells, 8 embryos; ***P <0.0001). (n) The average area of detected centrin-RFP foci is not significantly different between control and Rsg1 KD MCCs, indicating that there are no gross abnormalities in apically docked basal bodies upon Rsg1 KD (Ctl: 0.12 ? 0.02, n = 45 cells, 8 embryos vs. Rsg1 KD: 0.13 ? 0.03, n = 42 cells, 8 embryos; P = 0.685). Scale bars in a?f represent 5 ?m. Image published in: Brooks ER and Wallingford JB (2013) Copyright © 2013 Brooks and Wallingford; licensee BioMed Central Ltd. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution license Larger Image Printer Friendly View |