Fig. 3: Cervical cancer-produced NMB reprograms and activates Schwann cells.
From: Cervical cancer-produced neuromedin-B reprograms Schwann cells to initiate perineural invasion
a Phalloidin staining was analyzed in Schwann cells treated with supernatant from RSC96/HeLa/ME180 cells. Red, Phalloidin, F-actin staining. Blue, DAPI, Nuclei. Scale bars: 25 μm. b Immunofluorescent Nestin staining was analyzed in Schwann cells co-cultured with HeLa and ME180 cells. Scale bars: 500 μm. c cJUN and BDNF mRNA expression of cocultured RSC96 cells with cervical cancer cells were examined by qRT-PCR (n = 3, one-way ANOVA and Tukey’s multiple comparisons test). d EdU staining (red) was analyzed in RSC96 cells cocultured with HeLa and ME180 cells. Scale bars: 50 μm. e Cell migration and invasion of Schwann cells co-cultured with HeLa and ME180 cells were examined by Boyden chamber assay with or without Matrigel-coated inserts. Scale bars: 50 μm. f Phalloidin staining was analyzed in Schwann cells treated at gradient concentrations of NMB. Scale bars: 25 μm. g EdU staining was analyzed in RSC96 cells treated with NMB. Scale bars: 50 μm. h cJUN and BDNF mRNA expression of RSC96 cells upon treatment with different concentrations of NMB were examined by qRT-PCR (n = 3, one-way ANOVA and Tukey’s multiple comparisons test). i Cell migration and invasion assays of Schwann cells treated with NMB were examined by Boyden chamber assay with or without Matrigel-coated inserts. Scale bars: 50 μm. j Phalloidin staining was analyzed in Schwann cells treated with NMB knockout HeLa or ME180 cells. Scale bars: 25 μm. k EdU staining was analyzed in RSC96 cells treated with NMB knockout HeLa or ME180 cells. Scale bars: 50 μm. l cJUN and BDNF mRNA expression of cocultured RSC96 cells with NMB knockout cervical cancer cells were examined by qRT-PCR (n = 3, one-way ANOVA and Tukey’s multiple comparisons test). m Cell migration and invasion of Schwann cells treated with NMB knockout HeLa or ME180 cells were examined by Boyden chamber assay with or without Matrigel-coated inserts. Scale bars: 50 μm. Data are shown as photographs from one representative of three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.
