Fig. 4

Kaempferol and quercetin downregulate G9a and PAX3-FOXO1 in RMS cells. Rh30 (A) and Rh41 (B) cells were treated with kaempferol or quercetin for 24 h and whole cell lysates were analyzed by western blots as outlined in the Materials and methods. Rh30 (C) and Rh41 (D) cells were treated with kaempferol or quercetin for 24 h and G9a and PAX3-FOXO1 mRNA levels were determined by real time PCR as outlined in the Materials and methods. E Rh30 cells were treated with 50 μM kaempferol and quercetin for 24 h and analyzed in a ChIP assay and the PAX3-FOXO1 gene (F) was also normalized to IgG using the appropriate primers and RT-qPCR as outlined in the Materials and methods. These results are quantified in Supplemental Fig. 3. Results (C and D) are means ± SD for at least 3 determinations and significant (p < 0.05) inhibition is indicated (*)