Fig 6. Wnt5a stimulates expansion of B-1P cells in vitro.

B-1P cells were co-cultivated onto OP9-WT (WT), Wnt3a-transduced OP9 (Wnt3a) or Wnt5a-transduced OP9 (Wnt5a) layers. After 9 days, the number of B-1P cells in these cultures was analyzed. (A) Absolute number of B-1P cells co-cultivated with OP9-WT, OP9-Wnt3a and OP9-Wnt5a after 9 days of co-culture. The initial input of B-1P was 3x10³ cells, which is represented by red line. (B) Expansion of B-1 cell precursor population after 9 days in co-culture with OP9-WT, OP9-Wnt3a and OP9-Wnt5a. The graphs represents fold change increase in relation to input of cells. It was calculated as the number of B-1 cell precursors after 9 days in culture normalized by the initial input (3x10³ cells). (C) Dot plots of expression of CD19X AA4.1 by B-1P cells before culture and after 9 days in culture with OP9-WT, OP9-WNt3a, OP9-Wnt5a. Results are representative of 2 experiments performed in triplicate (n = 3). ***p<0,001, **p<0,01 and *p<0,05. (One way ANOVA).