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. 2016 Dec 21;7:13781. doi: 10.1038/ncomms13781

Figure 4. Molecular characterization of RAF1-deficient cells.

Figure 4

(a) siRNA-mediated RAF1 silencing promotes the proliferation of Hep3B (n=6), HuH-7 (n=5) and HepG2 (n=4) cells and increases the expression of YAP1 and GP130 as well as STAT3 phosphorylation. siRAF1#1 targets the region around nucleotide 721, while siRAF1#2 is a mixture of siRNAs targeting the region from nucleotide 692 to 1,093 in the RAF1 mRNA. (b) PCR and immunoblotting analysis of F/F and RAF1Δ/Δ DIH. (c) Morphology (x200 magnification) and molecular characterization (d) of primary hepatocytes (P-HEPS) compared to DIH (AFP, α-fetoprotein; ALB, albumin; TUBA, loading control). The immunoblot is representative of two independent experiments. (e) Proliferation of DIH in decreasing amounts of FBS. (f) Molecular defects of RAF1-deficient P-HEPS and DIH treated with the indicated concentrations of IL6 for 30 min. TUBA serves as loading control. Data are presented as mean±s.e.m. *P≤0.05, **P<0.01, ***P<0.005 according to Student's t test. See also Supplementary Fig. 5.