Fig. 1: Hedgehog responsiveness and primary cilia formation in mouse osteosarcoma cells.
From: Ligand-dependent hedgehog signaling maintains an undifferentiated, malignant osteosarcoma phenotype
a Western blot analysis of SHH and ACTIN expression in radiation induced (45Ca) and genetic (Osx p53Rb KO) mouse osteosarcoma cell lines. Lysates from NIH-3T3 cells and recombinant SHH (rSHH) are included as positive controls. N = 5 independent cell lines per genotype. b Mutational status of Trp53, Rb1 and Cdkn2a in the 45Ca mouse osteosarcoma (CaOS) cell lines. c Gli1 mRNA expression in 45Ca and p53Rb KO mouse osteosarcoma cell lines treated with PBS vehicle control, 1 μg/ml rhSHH, 1 μg/ml rhSHH and 400 nM sonidegib (LDE225) or 400 nM sonidegib for 24 h. Values are normalized to the expression of β2-microglobulin. n = 3, mean ± SEM, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, one-way ANOVA/Tukey’s test. d Representative images demonstrating immunofluorescence colocalization of acetylated α-tubulin (AcTUB) and ARL13B in 45Ca (CaOS18, CaOS25) and p53Rb KO (C78F) mouse osteosarcoma cell lines cultured in 10% serum, or serum-free media for 24 h. Primary cilia are highlighted by arrows (scale bar, 5 µm). e Primary cilia frequency in 45Ca and Osx p53Rb KO mouse osteosarcoma cell lines cultured in 10% serum or serum-free media for 24 h. n = 5 individual replicates per cell line, mean ± SEM, *P < 0.05, **P < 0.01, ***P < 0.001, ****P ≤ 0.0001, Student’s unpaired t test.
