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. Author manuscript; available in PMC: 2012 Dec 9.

Published in final edited form as: Am J Transplant. 2009 Sep 25;9(12):2679–2696. doi: 10.1111/j.1600-6143.2009.02831.x

(A). Untreated ^WTC57BL/6 cultures show positive staining for ASK1p-Ser967 and ASK1p-Thr845 in CM (open arrows) and in VEC (arrows) (a,b). Untreated TNFR1^KO cultures are positive for ASK1p-Ser967 (arrowhead) but not for ASK1p-Thr845 (c,d). In contrast, TNF-treated ^WTC57BL/6 cultures show a diminished ASK1p-Ser967 (g) but a strong signal for ASK1p-Thr845 in CM (open arrow) and in VEC (arrowhead) (h) while TNF-treated TNFR1^KO are negative (i,j). In contrast, CM (open arrow) and VEC (arrow) are strongly positive for ASK1p-Thr845 but not for ASK1p-Ser967 in TNF-treated TNFR2^KO cultures (k–l). (B). A strong signal for total Etk and phosphorylated Etk (Etkp) is seen in CM (open arrow) and in VEC (arrow) in untreated ^WTC57BL/6 (a,b) and in TNFR1^KO cultures (c,d), while untreated TNFR2^KO cultures are negative (e,f). TNF-treated ^WTC57BL/6 and TNFR1^KO cultures show Etk and Etkp in VEC (arrows) and in CM (open arrows) (g–j). In contrast, TNF-treated TNFR2^KO cultures are negative (k,l). Images are representative of 3 independent experiments with similar results (Original Mags, ×63).

(A). Untreated ^WTC57BL/6 cultures show positive staining for ASK1p-Ser967 and ASK1p-Thr845 in CM (open arrows) and in VEC (arrows) (a,b). Untreated TNFR1^KO cultures are positive for ASK1p-Ser967 (arrowhead) but not for ASK1p-Thr845 (c,d). In contrast, TNF-treated ^WTC57BL/6 cultures show a diminished ASK1p-Ser967 (g) but a strong signal for ASK1p-Thr845 in CM (open arrow) and in VEC (arrowhead) (h) while TNF-treated TNFR1^KO are negative (i,j). In contrast, CM (open arrow) and VEC (arrow) are strongly positive for ASK1p-Thr845 but not for ASK1p-Ser967 in TNF-treated TNFR2^KO cultures (k–l). (B). A strong signal for total Etk and phosphorylated Etk (Etkp) is seen in CM (open arrow) and in VEC (arrow) in untreated ^WTC57BL/6 (a,b) and in TNFR1^KO cultures (c,d), while untreated TNFR2^KO cultures are negative (e,f). TNF-treated ^WTC57BL/6 and TNFR1^KO cultures show Etk and Etkp in VEC (arrows) and in CM (open arrows) (g–j). In contrast, TNF-treated TNFR2^KO cultures are negative (k,l). Images are representative of 3 independent experiments with similar results (Original Mags, ×63).