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. 2012 Sep 4;122(10):3603–3617. doi: 10.1172/JCI62229

Figure 4. HOXA9 expression in EOC cells induces CAF features in normal omental fibroblasts.

Figure 4

(A) Diagram of fibroblast-priming assays. +HOXA9 control and HOXA9-knockdown SKOV3ip cells were cultured for 2 days to generate tumor-conditioned media (shown in pink). Normal omental fibroblasts were incubated for 5 days in SKOV3ip-conditioned medium (i.e., primed) or nonconditioned medium (i.e., unprimed) and then analyzed by Western blot and qRT-PCR. Fresh nonconditioned medium was added to washed fibroblasts. At 2 days thereafter, medium conditioned by fibroblasts (shown in light blue) was analyzed by ELISA and used for incubating control (nontargeting) SKOV3ip cells. (B) Western blot of αSMA levels in unprimed and primed fibroblasts. Full uncut gels are shown in the Supplemental Material. (C) Levels of growth factors in media conditioned by unprimed and primed fibroblasts. (D) Growth rates of control SKOV3ip cells incubated in fibroblast-conditioned medium. Average results of assays using 3 independent sets of each type of fibroblast-conditioned medium are shown in C and D. (E) Relative growth of control SKOV3ip cells at 6 days after incubation in medium conditioned by fibroblasts that were initially primed in +HOXA9 control SKOV3ip-conditioned medium, where fibroblast-conditioned medium was left untreated, treated with IgG, or depleted by IP with Abs to CXCL12 and IL-6. *P < 0.001; #P < 0.005.