(A) 3D co-cultures of ECs (red) and KRC PCCs (green) shows that compared with vehicle (DMSO, left), ruxolitinib ([100 nM], right) suppresses PCC growth. Shown are representative phase contrast and fluorescent images taken on day 8. Scale bars, 200 μm. (B) Fluorescence intensity quantitation shows that compared with 3D cultures in which ECs and PCCs are cultured independently (single culture), culturing ECs and PCCs together in 3D (co-culture) significantly enhances PCC growth, which is blocked by ruxolitinib (open bars). Data are mean ± SEM from three independent experiments. *P < 0.05, and **P < 0.01. (C) Schematic representation of PCC and EC cross-talk. TGF-β activates canonical Smad-dependent signaling in PCCs (top) leading to enhanced production of pro-angiogenic factors, which can be blocked by SB505124. These factors activate JAK/STAT3 signaling in ECs (bottom), which promotes EC proliferation through HDAC9, and ruxolitinib blocks these effects. ECs also produce factors (angiocrine factors) that can exert growth-stimulatory effects on PCCs through JAK/STAT3 signaling, which can also be targeted with ruxolitinib.