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. 2015 Nov 9;5:16513. doi: 10.1038/srep16513

Figure 2. Effects of Shh signaling on Stil expression and PC12 cell survival in response to toxic drug treatment.

Figure 2

(A) Western blot of STIL proteins in PC12 cells after transfection with pSIREN-Stil-shRNA (knockdown) or pCS2+Flh-Stil plasmid (overexpression). The gels have been run under the same experimental conditions. (B,C) Western blot of STIL and GLI1 proteins in PC12 cells after treatment with Smo receptor agonist cyclopamine or Smo receptor antagonist purmorphamine. Treatment with cyclopamine decreased GLI1 expression but resulted in the accumulation of STIL proteins. Treatment with purmorphamine promoted Shh signaling but caused a decrease in STIL protein expression. All the gels have been run under the same experimental conditions. (D) RT-PCR analyses of Stil mRNA expression in PC12 cells transfected with pSIREN-Stil-shRNA or pCS2+Flh-Stil plasmid. Note the alterations in Stil mRNA expression in response to plasmid transfection. (E,F) RT-PCR analyses of Stil and Gli1 expression in response to cyclopamine or purmorphamine treatment in PC12 cells. Treatment with either compound resulted in significant changes in the expression of Stil or Gli1 mRNA. (G,H) Cell survival in response to 6-OHDA treatment (75 μM) in proliferating and NGF-induced mature cells while the Shh signaling transduction was modified by plasmid transfection. Data represent the Means ± SE, n = 4; n.s., not significant, *p < 0.01.