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. 2023 Jul 27;25(12):2177–2190. doi: 10.1093/neuonc/noad134

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© The Author(s) 2023. Published by Oxford University Press on behalf of the Society for Neuro-Oncology. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com

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Figure 3. — Fatty acid-binding protein 7 (FABP7) drives xenograft GBM growth and metastasis. (A) Real-time dynamic changes incell density in the scratch as a function of migration start time (Incucyte). (B) Real-time dynamic changes in the percentage of GFP intensity within the closing gap (Incucyte). (C, D) Tumor volume as a function of days post injection of U87-pREP4 and U87-pREP4-FABP7 cells in the flank of nude mice (C) and statistical analysis of tumor volume (D) at specific time points (inset shows photograms of tumors). (E) Representative images showing FABP7 immunostaining in xenograft tumors generated from FABP7-expressing (pREP4-FABP7; left, middle – magnification of the squared area of the left panel) and control (pREP4) U87 cells (right panel). (F) Western blot showing expression of FABP7 and SOX2 in primary cultures of xenograft tumor tissues, as well as in intestinal metastasis cultured in the absence and presence of hygromycin. (G) Representative images of Transwell assay comparing migration of the intestinal metastasis-derived cells without (Met) or with hygromycin selection (Met + hygro). (H) Histogram showing quantification and statistical significance of the results obtained using the Transwell assay. (I-K) Representative images of tumors formed from U87 pREP4 (I) and U87-pREP4-FABP7 (J) cells orthotopically injected into the brains of NSG mice. Magnified images (K) show the morphology of FABP7-positive cells from the indicated areas in M with arrowheads indicating extended cell processes. Large arrows in I and J point to tumors. Small arrows in J point to FABP7 immunoreactive cells. (L) Representative images showing the distribution of A4-007 FABP7-positive cells orthotopically injected into the brains of NSG mice (left panel), and the effect of intraperitoneal injection of the FABP7 inhibitor SBFI-26 on tumor formation (right panel). The flank xenograft experiments were performed twice with 6 mice in each group per experiment and the two orthotopic xenograft experiments were each carried out once with 3 mice in each group. P denotes pseudopalisade. N, necrosis. **, P < .01. ***, P < .001. ****, P < .0001.