Figure 2.

Effects of ADAM10 inhibitors on ATP content, lactate dehydrogenase or TNFa release and size of autologous lymph node-mesenchymal stromal/Hodgkin lymphoma cell spheroids. Autologous mixed spheroids of LN-MSC773 (2x10⁵) and RS773 cells (4x10⁵) were prepared as previously described.³¹ (A to C) 10 mM (panel Ai) or dilutions (panel Aii), B and C) of LT4 or MN8 or GIX were added to the mixed spheroids for 48 hours (h) (Ai) and C), 72 h (Ai)) or 96 h (Ai), Aii) and B). At the indicated time points ATP (Ai) and Aii)), lactate dehydrogenase (LDH) (B) or TNFa (C) were measured by specific assays. Results are expressed as luminescence arbitrary units (a.u./10⁴cells, A) or O.D.490 (a.u./10mcells, B) or pg/mL/10⁴cells (C) and are referred to one representative experiment out of three in triplicate. (D) Spheroid area was measured as previously described³¹ in each culture well at 48 h (left), 72 h (central), 96 h (right). At least triplicates were analyzed for each culture condition and a minimum of 50 single spheroids for each of three independent experiment. Nil: no drug added (medium with dimethyl sulfoxide 1:1,000). Mean ± standard deviation from three experiments is indicated. *P<0.01 and **P<0.001 vs. nil.