Figure 2.
PAD3 is up-regulated with development and following injury at E15. A) PAD mRNA detected by qPCR shows almost 10 fold increase 24 hours after injury at E15; note also increased PAD3 in sham-operated spinal cords between E15 and E16. *p<0.05. B) Western blot with an anti-PAD3 antibody of total proteins from E11 spinal cords and reactivity of anti-PAD3 and anti-PAD2 antibodies in nuclear (N) and cytoplasmic (C) fractions. C) E12 spinal cord double-labelled for PAD3 (red) and NeuN (green). D) Central canal region in an E12 spinal cord stained for PAD3 (red) and counterstained with the nuclear stain Hoechst dye (blue); note the largely nuclear localization of PAD3. E) High magnification of the gray matter (region boxed in C) showing some neurones double-labelled for PAD3 (red) and NeuN (green). F) PAD3 expression in E15 spinal cord; note PAD3 expression both in gray and white matter. G) Cells double-labelled for the oligodendrocyte marker MBP (green) and PAD3 (red) are present in the white matter of E16 spinal cords (white arrow); PAD3-positive/MBP-negative cells are also observed (yellow arrowhead). Scale bars = 200 μm (C,F); 40 μm (D); 10 μm (E, G).