Fig. 3

SMO-193a.a. maintains self-renewal of CSCs. a IB of SMO-193a.a. in 456, 3691 CSCs stably knocking down (KD) circ-SMO and re-expressed circ-SMO or SMO-193a.a. linearized vector. b qPCR of circ-SMO in in 456, 3691 CSCs KD circ-SMO and re-expressed circ-SMO or SMO-193a.a. linearized vector. c Limited dilution assay (LDA) of 456, 3691 CSCs with indicated modifications. Stem cell frequencies were calculated. Lines show the estimate values, upper/lower limits of confidence intervals. d Cell proliferation of 456, 3691 CSCs with indicated modifications. e EdU incorporation assay of 456, 3691 CSCs with indicated modifications. f Stemness markers including Sox2, Oct4, Nestin, and Nanog; differentiation markers GFAP and Tuj-1 were determined by IB in 456, 3691 CSCs with indicated modifications. g Upper, Illustration of circ-SMO mutant vector. An A was inserted after the start codon of SMO-193a.a. to compromise the ORF. Lower, qPCR and IB were used to verify circ-SMO mutant vector. h LDA of 456, 3691 CSCs with indicated modifications. Stem cell frequencies were calculated. Lines show the estimate values, upper/lower limits of confidence intervals. i Cell proliferation of 456, 3691 CSCs with indicated modifications. j EdU incorporation assay of 456, 3691 CSCs with indicated modifications. Lines show the mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001. In a to j, data are representative from at least three experiments with similar results