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. 2012 Sep 28;3(10):1169–1181. doi: 10.18632/oncotarget.600

Figure 4. JIP1-inhibitor BI-78D3 reduces JIP1/p-JNK complexes in doxorubicin treated OS cells.

Figure 4

(A) Western blot analysis of baseline JIP1 levels in human OS cell lines and human primary osteoblast culture Hum71. All OS cell lines exhibit higher JIP1 levels than the human primary osteoblasts. (B) Immunoprecipitation and Western blot analysis of the JIP1/JNK activation module. LM7 and U2OS cells were treated with doxorubicin either in the presence or absence of BI-78D3. Whole cell lysates were analysed for JIP1 expression with β-actin serving as control for equal loading. In addition, cell lysates were immunoprecipitated with an anti-JIP1 antibody and Western blot analysis was done with an anti-p-JNK antibody. Doxorubicin-treated LM7 cells show an increased expression level of p-JNK compared to untreated cells, whereas in U2OS there is a slight decrease in p-JNK after doxorubicin treatment. After concurrent treatment with BI-78D3, p-JNK expression levels are diminished, indicative of inhibition of JIP1 scaffold function.