Skip to main content
. Author manuscript; available in PMC: 2017 Feb 10.
Published in final edited form as: Oncogene. 2016 Jul 18;36(6):850–862. doi: 10.1038/onc.2016.254

Figure 4. FoxF1 or FoxF2 knockdown alters expression of p21Cip1 /p27Kip1 pathway-related genes.

Figure 4

Total RNA and protein from orthotopicaly-grown RMS tumors were harvested 3 weeks after 76-9 tumor cells inoculation. (A) Depletion of FoxF1 or FoxF2 (FoxF1-KD, FoxF2-KD) in RMS tumor cells decreased mRNAs of cyclin D1, cyclin A2, cyclin E and C-myc, all of which are G1/S-promoting cell cycle genes. p21 mRNA was increased in the FoxF1- and FoxF2-depleted RMS tumors. qRT-PCR was used to assess expression levels. β-actin mRNA was used for normalization. A p value <0.05 is shown with (*). (B) No changes in mRNAs of G2/M cell cycle transition and cytokinesis genes, Cdc25b, cyclin B1, Plk1 and Aurora B, were found using qRT-PCR. β-actin mRNA was used for normalization. (C) Western blot demonstrated increased expression of p21Cip1, p27Kip1 and pRb (Ser807/811) proteins in FoxF1-KD or FoxF2-KD RMS tumors compared to control. (D) Western blot showed decreased protein levels of Cdk1, Cdk2, Cdk4, Cdk6, cyclin D1, and cyclin E1 in FoxF1-KD or FoxF2-KD RMS tumors compared to control. β-Actin was used as loading control. (E) qRT-PCR was performed using human rhabdomyosarcoma Rh30 cells with stable knockdown of FoxF1 or FoxF2. β-actin mRNA was used for normalization.