Loss of prx1 expression is associated with a compensatory
increase in Prx5. A, total cell lysates from
prx1+/+ and prx1-/- MEFs were
immunoblotted and probed with antibodies against the indicated Prx family
members or against β-tubulin as a loading control. B,
quantitative real time PCR analyses of transcript levels for individual
members of the peroxiredoxin family. Samples were run in triplicate and
expressed as a mean relative expression ratio when compared with
glyceraldehyde-3-phosphate dehydrogenase ± S.E. *,
p value was significant (p = 0.012). C, Prx5
immunoblots of prx1-/- MEFs stably transduced with a
lentiviral-prx1 vector or control vector. Note that the reexpression
of Prx1 was associated with a concurrent reduction/normalization of Prx5.
D, immunostaining for prx5 in prx1+/+
and prx1-/- MEFs. A FITC-tagged secondary antibody was
used for visualization. The bottom panel shows the same cells
counterstained with 4′,6-diamidino-2-phenylindole to emphasize the
nucleus. Note the stronger signal in prx1-/- MEFs, which
is mostly diffuse cytoplasmic staining. Some cells display punctate
cytoplasmic staining, which is shown by the arrow in the
inset of the top left panel.