Fig 2: Measurement of Gli1 and MGMT expression by real-time quantitative PCR (a) and western blot analysis (b) in the three GBM lines, including A172, U251, and U87. The expression of the ubiquitously expressed β-actin gene was measured as the control. The MGMT promoter methylation status was also as showed (c). The lane labelled as ‘M’ means methylated; ‘U’ stands for unmethylated. Gli1 expression was increased by pcDNA3.1-Gli1 transfection in A172 cells measured at mRNA and protein levels since 48 h and thereafter (d–f), as well as MGMT expression and promoter status. When exposed to TMZ treatment, cell viability in the A172 cells transfected with pcDNA3.1-Gli1 was significantly higher than that in the control since 96 h and thereafter (g). The percentage of apoptotic cells at 120 h after pcDNA3.1-Gli1 transfection was significantly lower than in the control cells (h). *p < 0.05; **p < 0.01

Fig 3: The mRNA and protein levels of Gli1 and MGMT in U251 cells were significantly inhibited 48 h after treated by 5 μM cyclopamine (a and b). The MGMT promoter statement remained unchanged like the A172 treatment (c). When exposed to TMZ, cell viability treated with cyclopamine was significantly lower than that in the control 72 h after the treatment (d). The percentage of apoptotic cells in U251 cells treated with cyclopamine was significantly higher than in the control at 120 h post-treatment (e)

Fig 4: Relationship between MGMT expression, Gli1 nuclear staining and MGMT promoter methylation status in 48 cases of surgically-resected primary GBM tissues (a, b). Representative photomicrographs showed two GBM cases with distinct Gli1 activity (c). The top two photos represent a Gli1-positive GBM with visible nuclear staining of Gli1 (left) and the same sample stained with MGMT antibody showed that the percentage of MGMT staining was far more than 5% (right). The bottom two photos represent a Gli1-negative case with invisible nuclear staining of Gli1 (left). The same sample stained with MGMT antibody showed the ratio of MGMT staining was less than 5% (200×). The scale bar on the top left panel applies to the rest of the photographs and indicates 20 μm

Fig 5: Secretion of FGF21 by hepatocytes. FGF21 was quantified in (A) serum and (B) hepatocyte supernatants from male Hh-WT and Hh-KO mice by ELISA. Data are plotted as the mean ± standard deviation, n = 3–8, unpaired t-tests with p-values * ≤ 0.05 and *** ≤ 0.001. (C) Binding sites of GLI1 in potential regulators of Fgf21 expression identified by ChIP-seq. Numerical values are available in Table S9.