Fig. 5: NOTCH activation leads to reduced multiciliation in CP tumors.
A Immunofluorescent staining for ARL13B (yellow) is shown at day E14.5 in the upper roof plate progenitors (marked by dotted lines and orange arrowheads) and the CP epithelial cells (arrows) in the hindbrain in wild type and Lcre;Ptchcko animals, and CPP and abnormal CP growth (white arrowheads and dotted lines) in Lcre;NICD1 and Lcre;Ptchcko;NICD1 animals, respectively. GFP (green) labels tumor cells. DAPI staining (cyan) labels nuclei. Scale bar, 10 µm. Results were obtained from at least three independent experiments. The expression of ARL13B (red) is shown in tumor cells infected with viruses expressing GFP-tagged dnMAML1 or GFP (B), or treated with vehicle, or IMR-1/IMR-1A (C). GFP (green) labels infected or treated cells. DAPI staining (blue) labels nuclei. Scale bars, 20 µm. The percentage of multiciliated tumor cells after treatment is shown (n = 3, mean ± s.e.m., two-tailed unpaired t-test, **P < 0.01, ***P < 0.001). Results were obtained from at least three independent experiments, respectively. D The expression of Ki-67 (red) is shown in GFP+ tumor cells treated with vehicle or IMR-1. Quantitation of Ki-67 expression is shown (n = 6 per treatment; mean ± s.e.m., paired t-test, ***P < 0.001). Results were obtained from at least three independent experiments. E RNAscope analysis of Foxj1 expression (red) is shown in tumor cells treated with vehicle or IMR-1. DAPI staining (blue) labels nuclei. Scale bar, 20 µm. Quantification of Foxj1 transcript is shown on the right (n = 7 per treatment; mean ± s.e.m., two-tailed unpaired t-test, **P < 0.01). Data are representative of three independent experiments.
