Figure 3.
Chromosomal aberration assays. (A) Near-diploid HCT 116 colorectal carcinoma cells were transfected with indicated miRNA plasmids to create stable cell lines. Cell clones were isolated, expanded and characterized for p54(nrb) expression by immunoblotting as described in ‘Material and Methods’ section. Replicate immunoblots were probed with anti-p54(nrb) and anti-GAPDH as an internal control. (B) Metaphase spreads were prepared and analyzed from irradiated p54(nrb)-deficient HCT 116 cell line as described in ‘Materials and Methods’ section. A representative image is shown. (C) Summary of chromosomal aberration data. Fifty metaphases were analyzed for each clonal cell line. The two clonal cell lines of each type (control and p54(nrb)-deficient) gave very similar results and the data were pooled for analysis. P-values were determined using a two-tailed Fisher’s exact test.