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Fig. 4 | Journal of Experimental & Clinical Cancer Research

Fig. 4

From: Autophagy-based unconventional secretion of HMGB1 in glioblastoma promotes chemosensitivity to temozolomide through macrophage M1-like polarization

Fig. 4

HMGB1 enhances the sensitivity of TMZ therapy by inducing M1-like polarization of TAMs. A-B Bioluminescent images (A) and quantification of tumors in mice (B) implanted with GL261 cells with treatment of Ctrl, TMZ, rmHMGB1 and TMZ plus rmHMGB1 for 7, 14 and 21 days. C The survival curves of tumor-bearing mice implanted with GL261 cells for indicated treatments. n = 8. D Representative flow cytometric analysis of tumor-infiltrating CD11c+ TAMs gated on live CD45+ CD11b+ TAMs isolated from GL261 cell-derived xenograft tumors with treatment of Ctrl (C), TMZ (T), rmHMGB1 (H) and TMZ plus rmHMGB1 (T + H) for 21 days (left panel). The histogram showed the statistical analysis (right panel). n = 4. E Representative flow cytometric analysis of tumor-infiltrating CD206+ TAMs gated on live CD45+ CD11b+ TAMs in GL261 cell-derived xenograft tumors with treatment of Ctrl (C), TMZ (T), rmHMGB1 (H) and TMZ plus rmHMGB1 (T + H) for 21 days (left panel). The histogram showed the statistical analysis (right panel). n = 4. F Representative IF of HMGB1 (purple), pan macrophage marker IBA1 (red) and M1-like TAM marker CD16/32 (green) in GL261 cell-derived xenograft tumors treated with TMZ plus rmHMGB1 (left panel). Quantitation of IBA1+/CD16/32+ TAM population in Ctrl, TMZ, rmHMGB1 and TMZ plus rmHMGB1 treatment groups (right panel). Scale bars = 50 μm. G Representative IF of HMGB1 (purple), IBA1 (red) and M2-like TAM marker CD206 (green) in GL261 cell-derived xenograft tumors treated with TMZ plus rmHMGB1 (left panel). Quantitation of IBA1+/CD206+ TAM population in Ctrl, TMZ, rmHMGB1 and TMZ plus rmHMGB1 treatment groups (right panel). Scale bars = 50 μm. *P < 0.05, **P < 0.01, ***P < 0.001, ns = no significance

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