Fig. 1.

TWS119 and rapamycin induce phenotypic T_SCM cells. (a) Highly purified (T_SCM cell-freed) human nCD4 + T_N cells activated for 14 days in the presence of TWS119 (5 μM) or rapamycin (100 nM) form a small-sized lymphocyte population which is not induced in the absence of TWS119 or rapamycin (red arrows). n = 5. (b) The small-sized lymphocyte populations induced by TWS119 or rapamycin mainly consist of CCR7 +, CD45RA + T_N-like cells (gated on live CD3 +, CD4 + T cells) and exhibit a fraction of phenotypic CD95 +, CD58 + T_SCM cells with highest frequencies in the transition zone (TZ). The flow cytometry plot shows phenotypic T_SCM cells in the total T_N-like cell population. T_SCM cell frequencies are depicted as percentages of the defined zones. Data are represented as mean ± SEM. FSC = forward scatter. SSC = side scatter. RZ = rand zone. T_SCM = pharmacologically induced phenotypic T_SCM cells. n = 5. (c) Activation of highly purified human nCD4 + T_N cells for 14 days in the presence of PP242 (100 nM) or KU-0063794 (100 nM) also leads to the formation of the small-sized lymphocyte population (red arrows), containing phenotypic CD95 +, CD58 + T_SCM cells (shown in the total T_N-like cell population). In contrast, priming in the presence of the alternative Wnt activators Indirubin-3-monoxime (I-3-M, 4 μM) or Wnt3A (10 nM) does not lead to the formation of the small-sized lymphocyte population (red arrows). n = 4. T_SCM = pharmacologically induced phenotypic T_SCM cells.