Fig. 3.
Ectopic reconstitution of ING4 in T47D breast cancer cells. (A) Southern blot. Genomic DNA was digested with BamHI and analyzed by Southern blotting with 32P-labeled full length cDNA of ING4 or the actin gene, ACTB,as hybridization probes. (B) CGH analysis of T47D (filled circles) and MDAmb231 (open squares) cell lines. Loss and gain of DNA are reflected in the relative copy number score on the y axis of the graph. Relative copy numbers were determined by the ratio of the test DNA to the reference DNA. Therefore, a -0.5 score indicates that the test DNA contained half the copy number compared with the reference DNA, reflecting the loss of one copy in the test DNA. As such, loss of two copies would score -1.0. Small arrows mark the location of BAC probes, RP11-272L6 at 5.1 Mb and RP11-59H1 at 12.6 Mb from telomere. A cytological banding of chromosome 12, the location of ING4, and the extent of deletion in T47D are shown. (C) Suppression of growth in soft agar. T47D cells infected with the retroviral constructs, ING4-IRES-GFP and IRES-GFP, were plated in soft agar and then incubated for 21 days. Colonies were counted with the aid of either light or fluorescent microscopy.