Fig. 4: NK1R drives the development of NE phenotype and therapeutic resistance.

A The effect of NK1R overexpression on CgA, ENO2, AR, and PSA (left) and NE signature genes expression (right) in 22Rv1 and LNCaP. B Western blot analysis of NK1R expression in cells transfected by plenti-EGFP-Puro-CMV-NK1R (ov-NK1R) or control plasmid (Mock). Up: 22Rv1; down: LNCaP. GAPDH was used as control. C The effect of NK1R overexpression on colony formation, colonies were calculated with Image J (down). D The effect of NK1R overexpression on transwell cell invasion in 22Rv1 and LNCaP cells, the infiltrated cells were calculated with Image J (down). E The effect of NK1R activation by 1 μM hHK-1 on colony formation, colonies were calculated with Image J (right). F The effect of NK1R activation by 1 μM hHK-1 on transwell cell invasion in DU145/PC-3, 22Rv1/LNCaP, and 22Rv1-NE and LNCaP-NE cells, the infiltrated cells were calculated with Image J (right). G The effect of shRNA-mediated NK1R knockdown on CgA, ENO2 (left), and NE signature genes expression (right) in 22Rv1-NE and LNCaP-NE. H Tumor volume and weight of 22Rv1, 22Rv1-NE, and 22Rv1-NK1R xenografts grown in nude mice (n = 8). I Tumor volume and weight of 22Rv1-NE, 22Rv1-NE-shNK1R2#, DU145, and DU145-shNK1R2# xenografts grown in nude mice (n = 7). J IHC staining of NK1R, Ki67, CgA, ENO2 expression in 22Rv1/22Rv1-NK1R/22Rv1-NE xenografts. Scar bar, 50 μm. The intensity of relative protein was calculated with Image Pro Plus (down). K IHC staining of NK1R, Ki67, CgA, ENO2 expression in 22Rv1-NE/22Rv1-NE-shNK1R2# (up) and DU145/DU145-shNK1R2# xenografts (down). Scar bar, 50 μm. The intensity of relative protein was calculated with Image Pro Plus (right). Data were shown as mean ± SD, *P < 0.05, **P < 0.01, ***P < 0.001.