Journal: International Journal of Molecular Sciences
Article Title: Notch–Sox9 Axis Mediates Hepatocyte Dedifferentiation in Kras G12V -Induced Zebrafish Hepatocellular Carcinoma
Figure Lengend Snippet: The Notch signaling pathway was upregulated after hepatocellular carcinoma induction, and the inhibition of Notch signaling suppressed hepatocyte dedifferentiation. ( A ) WISH results showed the expression pattern of notch1a , notch1b , notch2 , notch3 , and her15 at 10 dpf after DOX activation of kras G12V . ( B ) Tg(fabp10a:Tet3G;TRE3G:kras G12V -ZsGreen1) with Tg(Hsp70l:dnMAML-GFP) double transgenic fish line treatment strategy. ( C ) Monolayer images of Anxa4 antibody staining showed expression changes in the DOX-induced kras G12V + and kras G12V + and dnMAML+ groups at 8 dpf and 10 dpf. ( D ) Statistics of Anxa4 + and ZsGreen1 + /ZsGreen1+ ratio in the DOX-induced kras G12V + ( n = 11) and kras G12V + and dnMAML+ ( n = 13) groups at 8 dpf and 10 dpf. ( E ) The results of fluorescent in situ hybridization (FISH) showed sox9b expression in the DOX-induced control and kras G12V + and kras G12V + and dnMAML+ groups at 10 dpf. ( F ) The results of FISH showed cp expression in the DOX-induced control, kras G12V + and kras G12V + and dnMAML+ groups at 10 dpf. Numbers indicate the proportion of larvae exhibiting that expression. Asterisks show significance: ****— p < 0.0001. Scale bars—100 μm; error bars—S.D.
Article Snippet: The cDNA of full-length Zebrafish kras was amplified by PrimeSTAR HS DNA polymerase (Takara) and point mutated to obtain kras G12V , cloned into the modified pTRE3G-BI:ZsGreen1 (Cat.631342, Clontech) vector; and TRE3G-BI is a bidirectional version of TRE3G promoter that allows for simultaneous, equivalent, and inducible expression of two transgenes.
Techniques: Inhibition, Expressing, Activation Assay, Transgenic Assay, Staining, In Situ Hybridization