Fig. 4: NMB receptor in Schwann cells mediates PNI of cervical cancer.

a Immunofluorescent GFAP (red) and NMBR (green) staining were analyzed in sciatic nerve PNI model conducted with HeLa (middle) and ME180 cells (lower). Scale bars: 7.5 μm. b, c Pearson colocalization coefficient (Rr) analysis of GFAP and NMBR in (a) HeLa co-cultured group (b) and ME180 co-cultured group (c) were shown. d Immunofluorescent NMB (green) and NMBR (red) staining were analyzed in Schwann cells co-cultured with HeLa cells. e, f Multicolor fluorescence staining was analyzed in specimens of cervical cancer injected mice (e) and cervical cancer patients with PNI (f). Green: Pan-CK, cyan: NMB, purple: GFAP, red: NMBR. Scale bars: 10 μm. g Representative images of mice three days after tumor injection revealing left hind limb function in the normal PBS, HeLa/ME180 with or without PD168368 groups (n = 5). h Quantification of SFI (paw span in millimeters) in PNI mice treated with or without PD168368 at three days, one, two, and five weeks after tumor injection was shown (n = 5). i Quantification of SFI in PNI mice treated with or without PD168368 at day three after tumor implantation was shown (n = 5, one-way ANOVA and Tukey’s multiple comparisons test). j Quantification of sciatic nerve function scores in PNI mice treated with or without PD168368 at three days, one, two, and five weeks after tumor implantation was shown (n = 5). k Quantification of sciatic nerve function scores in PNI mice treated with or without PD168368 at day three after tumor implantation was shown (n = 5, one-way ANOVA and Tukey’s multiple comparisons test). l Quantification of the length of PNI as measured by ultrasound imaging in PNI mice treated with or without PD168368 was shown (n = 5, one-way ANOVA and Tukey’s multiple comparisons test). m Quantification of tumor weight in PNI mice treated with or without PD168368 at day three after tumor implantation was shown (n = 5, one-way ANOVA and Tukey’s multiple comparisons test). n Neurons were analyzed by PGP9.5 staining in ME180 subcutaneous tumor-bearing mice treated with NMBR antagonist PD168368. scale bars: left,1 mm; right, 100 μm. o Quantification of intra-cervical cancer nerve numbers in ME180 subcutaneous tumor-bearing mice treated with NMBR antagonist PD168368 was shown (n = 5, unpair t-test). Data are shown as photographs from one representative of three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.