Fig. 3: SALL4 positively regulates the SHH pathway.

The transcriptional activity of A endogenous and B. C exogenous GLI1 has been assessed in the presence of ectopic SALL4 in HEK293Ts transiently transfected with a Firefly luciferase reporter gene under the control of a synthetic promoter containing 12 binding sites for GLI1 (12 × GLI1-BS), pRL-TK Renilla as normalization control, and the indicated plasmids. D GLI1-mediated transcription has been evaluated in HEK293Ts transiently transfected with constructs expressing Firefly luciferase under the control of PTCH promoter presenting a conserved (P1A WT-Luc) or mutated (P1A Mut-Luc) GLI1-BS, pRL-TK Renilla as normalization control, and the indicated plasmids. Luciferase analyses were performed 24 h after transfection. Data in A–C represent the mean of n = 3 independent experiments ± SD; data in D represent the mean of n = 4 independent experiments ± SD. All data are expressed as FC versus empty vector. Representative immunoblotting of n = 3 biological replicas with similar results showing the expression levels of transfected plasmids are reported. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001 calculated with two-sided Student’s t-test.