Figure 7

Phosphorylation of JAK downstream targets in cells from lymph nodes draining inflamed and noninflamed paws. Popliteal lymph nodes (i, ipsilateral; c, contralateral) were dissected from two animals each at 1 and 2 h post i.pl. CFA injection: Cells were isolated and cell lysates were analyzed using Western Blotting. Densitometry results are given as mean % expression of loading control ± SEM in the bar graphs. A) Tyrosine 641-phosphorylation of STAT6 (110 kDa) in CFA- and IL-4-stimulated lymphocytes, two out of 6 nodes analyzed per time point are shown. A commercially available phospho-STAT6 positive control (POS, BioLabs) was run on the same gel. M = protein marker. B) Tyrosine 705 (79/86 kDa)- and Serine 727 (86 kDa)-phosphorylation of STAT3, non-phosphorylated STAT3 (79 kDa), phosphorylated STAT1 (84/91 kDa), non-phosphorylated STAT1 (84/91 kDa), Threonine 308-phosphorylated Akt (60 kDa), and non-phosphorylated Akt (60 kDa). Stainings were performed on 3 nodes per time point; the staining for Tyrosine-phosphorylated STAT3 was performed on 6 nodes per time point. Statistical comparison of the ipsi- and contralateral side was performed using the Wilcoxon Matched Pair test; *P < 0.05. Representative immunoblots are shown. The same blot was probed sequentially with different antibodies. Lanes shown were run on the same gel, noncontiguous lanes are separated by thin white lines. Ser, Serine; Thr, Threonine; Tyr, Tyrosine.