Figure 4.

HIPK2 depletion in mouse primary bone marrow cells. Bone marrow cells were explanted from long bones of sacrificed mice and were infected with recombinant lentiviruses carrying control (Ctr) or two different HIPK2‐interfering sequences (2496 or 913). Infected cells were selected in puromycine for 48 h and then were plated in methylcellulose in the presence of GM‐CSF. (a) Upper and middle panels: single colonies were picked‐up from methylcellulose and employed to extract RNA. Real‐time RT‐PCR was performed on indicated genes. Lower panel: mean ± standard deviation of number of colonies present in each plate and counted at the inverted microscope. (b) Left panels: indicative morphology of each single colony in methylcellulose. Right panel: single colonies were picked‐up from methylcellulose and their cells were cytocentrifugated and stained with May‐Grünwald–Giemsa stain to assess granulocyte and macrophage differentiation. ‘% TD cells’ indicates percentage of terminally differentiated cells.