WNT7A overexpression in K562-, BJAB- and CEM-Tet-WNT7A cells. Overexpression of WNT7A in these cell lines using the doxycycline-inducible lentiviral expression system was confirmed by qRT-PCR and Western blot assays. (A) Graphics showing the amplification curves obtained for WNT7A and RPL32 in cells treated or not with Doxycycline (Doxy) for 4 h. The WNT7A normalized relative ratio of the cells treated with Doxy was calculated utilizing the non-treated cells as calibrator and taking GAPDH, RPL32 and RPS18 as reference genes in two independent experiments. (B) Western blot assays (from 50 μg total protein) showing the presence of WNT7a protein after a 38-h incubation with Doxy. Photo-documentation of the Coomassie stained gel is included as protein loading control.