Figure 3

CCND1 amplification behaviour in bladder tumors. FISH identification of chromosome 11 centromere (green) and CCND1 gene (red) in paraffin-embedded tumors. DNA staining was performed with DAPI (blue). Black and white images correspond to DAPI reverse staining. (A-H) Metaphasic cells showing the proposed sequence of 11q13 amplicon fragmentation from HSRs to DMs. (I-K) Sample U-364 showed a complex pattern of CCND1 amplification. Three sub-populations were detected in this sample. (I) Sub-population with gain of whole chromosome 11. (J) Sub-population containing HSR with high-level amplification of CCND1 (K) Sub-population containing amplification of CCND1 and undetermined flanking material in HSR. (L-O) Peripheral location of DMs in metaphasic cells. (P-Q) CCND1-positive micronuclei, see arrows. In Q, note the elimination in the micronucleus of whole CCND1 copies, except those attached to the centromere. (R) Metaphasic cells containing a dicentric chromosome with two centromeric signals of chromosome 11 and CCND1 amplification, see asterisks. (S and T) CCND1 with HSRs appears to be forming internuclear bridges, see arrows. (U and V) Nuclear blebs as nuclear protrusions with high CCND1 signal.