Figure 5

Expression and functional characterization of SLC22A16 in DLBCL cells. (a) Expression of SLC22A16 mRNA levels was measured by quantitative real-time PCR in a panel of DLBCL tumors (n=10) and cell lines (n=5) as described in Materials and methods. (b) Analysis of OCI-LY7 cells expressing an empty vector control (EV) or HA-SLC22A16 was performed by western blot analysis with an anti-HA-specific antibody to confirm SLC22A16 expression (inset). Doxorubicin uptake was measured in OCI-LY7 EV (solid line, open triangle) or SLC22A16 (dashed line, square symbol) cells using ¹⁴C-doxorubicin as described in Materials and methods. The experiment shown is representative of three independent experiments and error bars for triplicate wells are shown. (c) Proliferation of OCI-LY7 EV (solid line) or SLC22A16 (dashed line) cells was measured in the presence of increasing doses of doxorubicin (0–10 μM) by ³H-TdR incorporation. Data from each cell line were normalized to its respective nil control and the experiment shown is representative of three independent experiments, error bars for triplicate wells are shown. (d) Bioinformatic modeling of SLC22A16 mutations shows their location in major facilitator superfamily domains, which were assigned by Pfam using a hidden Markov model. Tan regions are predicted to be trans-membrane helices and red residues (side chains shown) are the predicted pore. Each mutation site sits at the entrance/exit of the predicted translocation pore (yellow spheres).