Figure 4

Immunohistochemical analysis of NKCC1 phosphorylation. (A) A cryosection of a T9 DRG was co-stained with antisera raised against NKCC1 and p-NKCC1 after a 3-hr treatment with inflammatory mediators. The two antibodies showed an almost perfect co-staining. The calibration bar indicates 100 μm. (B) In contrast to the NKCC1 immunosignal, the p-NKCC1 signal increased already 1 hr after the start of the treatment. The change of p-NKCC1 immune fluorescence in treated DRGs (FI_test) compared to the respective contralateral control DRGs (FI_ctr) is expressed as ΔFI = (FI_test-FI_ctr)/FI_ctr·100 [%]. ΔFI (± SD; 3 animals) was 18 ± 5.9% (1 hr), 45 ± 3.5% (2 hr), and 54 ± 3.3% (3 hr). Indicated significance levels are p ≤ 0.05 (*) and p ≤ 0.001 (***). Numbers in parentheses indicate cells evaluated.