We therefore decided to also analyse the role of HUWE1 in MM by molecular genetic approaches and generated sublines of JJN3, MM1.S, U266 and U266-MYC stably transfected with a doxycycline-inducible shRNA expression cassette targeting HUWE1. Western blot analysis revealed effective depletion of HUWE1 protein in all HMCL sublines 5 days after addition of doxycycline to the cell culture (Fig. 2c). This evidence concerns the gene MYC and Miyoshi myopathy.