Fig 2: FDPS promotes tumour growth in a macrophage-dependent manner. A, The level of FDPS in GL261 cells was analysed by Western blotting. B and C, Representative immunohistochemistry of F4/80+ cells in sections from glioma tumours obtained from C57BL/6J mice treated with clodronate liposomes or PBS liposomes. D, Volume of GL261 tumours treated as indicated. Error bars represent the SD, *P < 0.05; **P < 0.01 (Scale bars, 50 µm)

Fig 3: CCL20 mediates the promoting role of macrophage infiltration induced by FDPS. A, The mRNA level of indicated chemokines in the conditional medium from U87 cells with or without FDPS silencing was analysed by real-time PCR. B, The mRNA level of CCL20 in the conditional medium from U251 cells with or without FDPS silencing was analysed by real-time PCR. C, The level of CCL20 in the conditional medium from U87 and U251 cells with or without FDPS silencing was analysed by ELISA. D, The level of CCL20 in the conditional medium from A172 and H4 cells with or without FDPS overexpression was analysed by ELISA. E, Volume of parental and FDPS-overexpressing GL261 tumours treated as indicated. F, F4/80+ cells in sections from indicated glioma tumours. Error bars represent the SD, *P < 0.05; **P < 0.01; ***P < 0.001

Fig 4: Immunolocalization of FPPS in the murine cochlea. (A) Low-magnification cross-section through the cochlea at 6 weeks revealed strong FPPS immunoreactivity (green) in the Myo7A-positive area of hair cells (white, arrow) and NF-H-positive SGNs (red). (B) High-magnification cross-section of the spiral ganglion co-immunostained for SGNs with NF-H (red) and FPPS (green) shows a weak positive signal in SGNs (white arrowhead), and a strong positive signal adjacent to SGNs where spiral ganglion Schwann or satellite glial cells reside (white arrow). (C) Low-magnification view of a neonatal cochlear explant at P4 reveals a strong FPPS signal (green) in the area of hair cells and the spiral ganglion. (D) High magnification of the spiral ganglion shows FPPS immunostaining (green) in (white arrowhead) and adjacent to (white arrow) SGNs (red). Scale bars: a, c, 100 µm; b, 50 µm; d, 20 µm. Representative images from N = 3 6-week-old mice and N = 3 4-day-old mice.

Fig 5: Wnt/ß-catenin signalling pathway is required for FDPS-mediated CCL20 induction. A, mRNA level of CCL20 in the conditional medium from A172 FDPS-overexpressing cells with or without XAV-939 pre-treatment was analysed by real-time PCR. B, mRNA level of CCL20 in the conditional medium from H4 FDPS-overexpressing cells with or without XAV-939 pre-treatment was analysed by real-time PCR. C, The level of CCL20 in the conditional medium from A172 FDPS-overexpressing cells with or without XAV-939 pre-treatment was analysed by ELISA. D, The level of CCL20 in the conditional medium from H4 FDPS-overexpressing cells with or without XAV-939 pre-treatment was analysed by ELISA. E, The level of CCL20 in the conditional medium from U87 FDPS knockdown cells with or without ß-catenin overexpression was analysed by ELISA. F, The level of CCL20 in the conditional medium from U251 FDPS knockdown cells with or without ß-catenin overexpression was analysed by ELISA. G, CCL20 was analysed by IHC in the shcon and shFDPS U87 tumours. H, CCL20 was analysed by IHC in the vector and FDPS GL261 tumours. I, p-ß-catenin was analysed by IHC in the shcon and shFDPS U87 tumours. J, p-ß-catenin was analysed by IHC in the vector and FDPS GL261 tumours. K, Volume of parental and FDPS-overexpressing GL261 tumours treated as indicated. L, F4/80+ cells in sections from indicated glioma tumours. Error bars represent the SD, *P < 0.05; **P < 0.01