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Fig. 2 INT6 is required for MEK protein levels and Erk-signaling. A. Osteosarcoma U2-OS cells untransfected (U), or transfected with siRNA targeted against the INT6 mRNA (si) or the reverse sequence (R), show reduced levels of INT6 and MEK1 protein specifically after transfection with INT6-siRNA, but no reduction in BAX, tubulin or actin protein levels. (B) Semi-quantitative-PCR shows MEK1 mRNA is unaffected in reverse sequence and INT6-siRNA treated cells, coupled with the expected reduced levels of the INT6 message in the INT6-siRNA transfected cells. c, PCR control without DNA. (C) Phospho-Erk levels are reduced in int6 morphants, while ponceau stain detects equal loading of protein on the gel.