Fig. 5

SMO-193a.a. interacts with SMO, promotes SMO activation, and is required for HH-induced SMO de-repression. a In vivo IP between SMO and SMO-193a.a. in 456 and 3691 CSCs. b In vitro IP using purified SMO and SMO-193a.a.. c Colocalization of SMO and SMO-193a.a.-mCherry in 387 CSC. Bars, 2 μm. d p-SMO and SMO were determined in 3691 and 387 CSCs with indicated modifications. e Upper left, Illustration of SMO-193a.a. 5 transmembrane helix structures and SMO 7 transmembrane helix structures. Upper right, Illustration of full-length and truncated SMO-HA-tagged plasmids. Lower, Full-length and truncated SMO as indicated were co-IP with SMO-193a.a. in 373 GBM cells with SMO-193a.a.-3XFlag and HA-SMO domains OV. f Upper, Illustration of cholesterol labeling experiments. A specific nucleotide sequence was labeled with cholesterol and added to 387 CSC with SMO-193a.a. OV or ctrl OV, or 3691 CSC with circ-SMO KD or ctrl KD. Lower, Cholesterol modified SMO or SMO-193a.a. were determined by IP followed qPCR. g 2.5μg/ml Shh was added to 456 and 3691 CSCs with indicated modifications. IB was used to determine the expression level of Gli1, p-SMO, and SMO at indicated timepoints. sh1 + 2 defines circ-SMO stable knocking down cells generated by using 1:1 cocktail mixture of sh1 and sh2. h Different concentrations of Shh was added to 456 and 3691 CSCs with indicated modifications. IB was used to determine the expression level of Gli1, p-SMO, and SMO after 24 h. Lines show the mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001. Data are representative from at least three experiments with similar results