Effects of BZ on hypoxia-selected K562 cells.

<p>Cells from day-7 hypoxic LC1 treated with BZ at day 6 as indicated were transferred into normoxic LC2 (3×10⁴ viable cells/ml) and trypan blue-negative cells counted at the indicated times of incubation in LC2. Values represent means±S.E.M. of data from 3 independent experiments.</p

Induction of apoptosis by BZ in hypoxia.

<p>(<b>A</b>) Total cell lysates in Laemmli buffer were subjected to immuno-blotting with the indicated antibodies. Anti-H4 was used to verify equalization of protein loading. One representative experiment out of 3 is shown. (<b>B</b>) Percentages cells undergone “early” (annexin-V+/PI-) or “late” (annexin-V+/PI+) apoptosis, as determined by flow-cytometry. Values are means±S.E.M. of 3 independent experiments.</p

Effects of BZ on hypoxia-resistant K562 cell subsets.

<p>Cells treated as indicated in hypoxic or normoxic LC1 (established at 3×10⁴ viable cells/ml) were transferred at day 2 (<b>A</b>) or day 7 (<b>B</b>) into normoxic LC2 (3×10⁴ viable cells/ml) and trypan blue-negative cells counted at the indicated times of incubation in LC2. Values represent means±S.E.M. of data from 3 independent experiments.</p

Effects of BZ on BCR/Abl protein expression in hypoxia.

<p>Total cell lysates in Laemmli buffer were subjected to immuno-blotting with an anti-Abl antibody. Anti-vinculin and anti-ERK1/2 antibodies were used to verify equalization of protein loading. One representative experiment out of 3 is shown.</p

Effects of Bortezomib (BZ) on K562 cell bulk.

<p>Cultures were treated or not with a single dose of 0.5 nM BZ from time 0 to day 7 or from day 1 to day 7 and trypan blue-negative cells counted at the indicated times. Values represent means±S.E.M. of data from 3 independent experiments. (<b>A</b>): hypoxia (∼0.1% O₂); (<b>B</b>): normoxia (21% O₂).</p