XB-IMG-133955
Xenbase Image ID: 133955
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Figure 3 GFP-IFT43 axonemal localization is reduced in Rsg1 KD MCCs. (a) High-magnification confocal image of axonemes from a control MCC expressing GFP-IFT43. The orange box indicates the region shown in a’. See also Additional file 3: Movie 3. (a’) A series of stills from Additional file 3: Movie 3. Yellow dashed lines outline the axoneme, and distal is to the right. Pink and blue arrowheads indicate an anterograde train and a retrograde train, respectively. (b) High-magnification confocal image of axonemes from a Rsg1 KD MCC expressing GFP-IFT43. Orange box represents the region shown in b’. See also Additional file 4: Movie 4. (b’) A series of stills from Additional file 4: Movie 4. Yellow dashed lines outline the axoneme, and distal is to the right. No IFT trains are visible during the course of the movie. Note that the entire axoneme exhibits a faint uniform background signal. (c–c”) A single confocal slice of a control MCC co-expressing membrane-RFP and GFP-IFT43. (d–d”) A single confocal slice of an Rsg1 KD MCC co-expressing membrane-RFP and GFP-IFT43. (e–e”) A signal confocal slice of an Rsg1 KD2 MCC co-expressing membrane-RFP and GFP-IFT43. (f) Quantification of mean GFP-IFT43 signal along control and Rsg1 KD axonemes, as normalized to membrane- RFP intensity along the same length. Note the decrease in normalized IFT43 signal in Rsg1 KD axonemes (Ctl: 0.28 ± 0.15, n = 225 axonemes, 28 cells, 5 embryos vs. Rsg1 KD: 0.15 ± 0.09, n = 250 axonemes, 32 cells, 6 embryos; ***P <0.0001). (g) Quantification of mean GFP-IFT43 signal along control and Rsg1 KD2 axonemes, as normalized to membrane-RFP intensity along the same length (Ctl: 0.26 ± 0.12, n = 200 axonemes, 44 cells, 6 embryos vs. Rsg1 KD: 0.15 ± 0.06, n = 223 axonemes, 52 cells, 6 embryos; ***P <0.0001). Scale bars in a–b’ indicate 3 μm. Scale bars in c–e” indicate 5 μm. Time stamps in a’ and b’ are relative to the first frame of Additional file 3: Movie 3 and Additional file 4: Movie 4, respectively. 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 |