Fig 6. Conditional deletion of calponin-3 does not affect early B cell development and function.

A. Percentages of different developmental stages and cell types derived from the bone marrow of control and B cell-specific Cnn3 knockout mice. Staining and gating of cells were performed according to Fig 4A. Control littermates (+/+ or +/f, positive for mb1-Cre) are depicted as black dots, knockout animals (f/d or f/f, positive for mb1-Cre) as white squares. Black bars mark the averaged percentage of cells for each subgroup. Percentages of cells in control and knockout animals were compared in an unpaired t-test (p>0.05 = not significant, n.s.). B. Western blot indicating induced signaling of control and calponin-3-deficient cells upon pre-BCR crosslinking. BM-derived B cells were starved for 30 min and then stimulated with an anti-μ antibody for 3 min. Cellular lysates were subjected to SDS-PAGE and western blotting. Anti-actin was used as a loading control, anti-calponin-3 confirmed the genotype of the used cultures. The band corresponding to calponin-3 is marked by an arrow, whereas non-specific signals from the polyclonal anti-calponin-3 antibody are labeled with asterisks. C. Induced calcium flux in B cell precursors derived from the bone marrow of control and knockout mice. Bone marrow cells cultured in the presence of IL-7 for 5 d were loaded with Indo-1, stimulated with pervanadate (marked by arrow) and analyzed by flow cytometry.