Figure 4. Loss of JNK impacts the IFT-B... | Rockefeller University Press

$Loss of JNK impacts the IFT-B complex and destabilizes the actin network of multiciliated cells. (A) Representative images from control and JNK morphant embryos injected with RFP-Centrin and GFP-IFT52. Scale bars, 10 µm. Quantification of the fluorescence intensity ratio of GFP-IFT52 shows impaired recruitment of IFT52 on basal bodies in JNK MO MCCs. Data represent mean ± SEM from three independent experiments. ****, P < 0.0001, unpaired two-tailed t test. (B) Immunofluorescence showing Xenopus epidermal MCCs labeled for F-actin (Phalloidin) in controls, embryos injected with JNK MO, or JNK MO/mScarlet JNK. Membrane-mCherry mRNA was used as a lineage tracer. Single optical sections at the level of apical and subapical actin layers show defective networks in JNK morphants. Scale bars, 10 µm. (C) Fluorescence recovery after photobleaching (FRAP) experiment on a stage 33 embryo with docked basal bodies, expressing either mKate2-actin alone or mKate2-actin with JNK MO. Normalized graph of FRAP experiment depicting signal intensity of bleached region over time shows faster recovery of apical actin and decreased immobile fraction in JNK MO (n = 8 MCCs from four embryos) MCCs compared with controls (n = 6 MCCs from three embryos). Error bars indicate ± SEM from two experiments. ***, P < 0.001, unpaired two-tailed t test. Scale bars, 10 µm. (D) Intercalating ciliated cells of stage 17 from control or JNK MO-injected embryos were stained for Phalloidin and centrin to label actin network and basal bodies, respectively. MCCs of JNK morphants show a defective association of basal bodies with the actin network. (E) Immunofluorescence image showing MCCs labeled for F-actin (Phalloidin) and pJNK. Scale bars, 10 µm. High-magnification top view and fluorescence intensity profile along the white line show that pJNK is interwoven within the apical actin network. Scale bars, 2 µm.$

Figure 4.

Loss of JNK impacts the IFT-B complex and destabilizes the actin network of multiciliated cells. (A) Representative images from control and JNK morphant embryos injected with RFP-Centrin and GFP-IFT52. Scale bars, 10 µm. Quantification of the fluorescence intensity ratio of GFP-IFT52 shows impaired recruitment of IFT52 on basal bodies in JNK MO MCCs. Data represent mean ± SEM from three independent experiments. ****, P < 0.0001, unpaired two-tailed t test. (B) Immunofluorescence showing Xenopus epidermal MCCs labeled for F-actin (Phalloidin) in controls, embryos injected with JNK MO, or JNK MO/mScarlet JNK. Membrane-mCherry mRNA was used as a lineage tracer. Single optical sections at the level of apical and subapical actin layers show defective networks in JNK morphants. Scale bars, 10 µm. (C) Fluorescence recovery after photobleaching (FRAP) experiment on a stage 33 embryo with docked basal bodies, expressing either mKate2-actin alone or mKate2-actin with JNK MO. Normalized graph of FRAP experiment depicting signal intensity of bleached region over time shows faster recovery of apical actin and decreased immobile fraction in JNK MO (n = 8 MCCs from four embryos) MCCs compared with controls (n = 6 MCCs from three embryos). Error bars indicate ± SEM from two experiments. ***, P < 0.001, unpaired two-tailed t test. Scale bars, 10 µm. (D) Intercalating ciliated cells of stage 17 from control or JNK MO-injected embryos were stained for Phalloidin and centrin to label actin network and basal bodies, respectively. MCCs of JNK morphants show a defective association of basal bodies with the actin network. (E) Immunofluorescence image showing MCCs labeled for F-actin (Phalloidin) and pJNK. Scale bars, 10 µm. High-magnification top view and fluorescence intensity profile along the white line show that pJNK is interwoven within the apical actin network. Scale bars, 2 µm.

This Feature Is Available To Subscribers Only