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Fig. 4 | Genome Biology

Fig. 4

From: A novel protein encoded by circular SMO RNA is essential for Hedgehog signaling activation and glioblastoma tumorigenicity

Fig. 4

SMO-193a.a. activates HH signaling in CSCs. a RNA-seq and Gene Set Enrichment Analysis (GSEA) of 456 and 3691 CSCs treated with circ-SMO scramble shRNA or sh1. b Gli1 binding site luciferase assay of 3691 and 387 CSCs with indicated modifications. c SMO and SMO-193a.a., SMO mRNA, and circ-SMO, as well as Gli1, C-Myc, and CCND1 mRNA expression were determined by IB or qPCR in 3691 CSC with indicated modifications. d SMO and SMO-193a.a., SMO mRNA, and circ-SMO, as well as Gli1, C-Myc, and CCND1 mRNA expression were determined by IB or qPCR in 387 CSC with indicated modifications. e IB of Gli1, c-Myc, and cyclin D1 expression in 3691, 387 CSCs with indicated modifications. f HH signaling agonist SAG and HH signaling inhibitor vismodegib were used in 3691 CSC with circ-SMO KD or in 387 CSC with circ-SMO OV. IB was used to determine the expression of indicated proteins. g Gli1 was OV in 456 and 3691 CSCs with circ-SMO KD. SMO-193a.a. and Gli1 level were determined by IB. h LDA was performed in 456 and 3691 CSCs with circ-SMO KD and Gli1 OV. Stem cell frequencies were calculated. Lines show the estimate values, upper/lower limits of confidence intervals. i Cell proliferation assay was performed in 456 and 3691 CSCs with circ-SMO KD and Gli1 OV. Lines show the mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001. In b to i, data are representative from at least three experiments with similar results

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