Fig. 2: Cervical cancer-produced NMB triggers PNI.

a mRNA levels of differential genes were analyzed using RNA-seq in tumor cells co-cultured with DRG. Quantification of NMB mRNA (b) and protein (c) in tumor cells co-cultured with RSC96 cells/DRG was shown (n = 3). d Quantification of NMB mRNA in specimens of cervical cancer patients (n = 306) and healthy women (n = 22) from TCGA and GTEx database was shown. e, f Immunohistochemistry (IHC) staining and relative IHC score of NMB were analyzed in normal (n = 8), CIN1 (n = 22), CIN2-3 (n = 22), CIS (n = 22) and cervical cancer (n = 38) tissues. ×25 magnification, scale bars: 1000 μm. g, h Immunohistochemistry (IHC) staining and relative IHC score of NMB were analyzed in patients with PNI (n = 20) and without PNI (n = 18). ×200 magnification, scale bars: 50 μm. i ROC curve for the sensitivity and specificity of NMB in detecting PNI by IHC staining. ROC, receiver operating characteristic. j Serum NMB was examined in healthy women (n = 8) and cervical cancer patients with (n = 20) or without (n = 18) PNI. k ROC curve for the sensitivity and specificity of NMB in serum tested by ELISA in detecting PNI. l Representative images of mice three days after tumor injection revealing left hind limb function in the normal PBS, sgNC, and sgNMB with/without recombinant NMB groups (n = 5). m Quantification of sciatic nerve function index (SFI) (paw span in millimeters) of mice in different groups at three days, one, two, and five weeks after tumor injection was shown (n = 5). n Quantification of SFI between different groups at day three after tumor implantation was shown (n = 5, one-way ANOVA and Tukey’s multiple comparisons test). o Quantification of the mean left sciatic nerve function scores of mice in different groups at three days, one, two, and five weeks after tumor injection was shown (n = 5). The score of 4 indicates normal function; score of 1 indicates total paralysis; a higher score indicates better sciatic nerve function. p Quantification of sciatic nerve function scores between different groups on day three after umor implantation was shown (n = 5, one-way ANOVA and Tukey’s multiple comparisons test). q Quantification of the length of PNI as measured by ultrasound imaging was shown (n = 5, one-way ANOVA and Tukey’s multiple comparisons test). r Quantification of tumor weight between different groups on day three after tumor implantation was shown (n = 5, one-way ANOVA and Tukey’s multiple comparisons test). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.