Fig. 8. Gata2-deficient ENC complemented with Gata2 exhibit much more active cyclin D1 and cyclin D2 transcription, while Gata2 transient overexpression blocks the proliferation of mouse neuroblastoma cells. (A-D) Gata2^–/– ENC were transfected either with pAdRSV-GATA2HA and pAdRSV-ßGal plasmids (A,C) or with pAdRSV-ßGal alone, then treated for X-Gal colouration and in situ hybridisation with cyclin D1 (A,B) or cyclin D2 (C,D) antisense RNA probes. (E-J) NB2a mouse neuroblastoma cells, either control (E-G) or transiently transfected with pAdRSV-GATA2HA (H-J). BrdU was added in the culture medium 30 hours after transfection, cells were recovered 15 hours later, processed for BrdU staining (F,I) and further HA-immunostained (I, red); nuclei were stained with DAPI (E,H) before mounting. (G,J) Superimpositions of E,F (G) and H,I (J). In the control culture (E-G), cells have not stopped growing and are all BrdU positive; whereas in transfected cultures (H-J), cells have not reached the same density and, especially GATA2-HA expressing cells, have not incorporated BrdU (J). Scale bar: 45 µm in A-D; 15 µm in E-J.