Fig 2: Representative images of immunofluorescence staining for TfR (green), NeuN (neuronal marker; red)/CD31 (vascular marker; red)/Olig2 (oligodendrocyte marker; red), and DAPI (nuclear marker; blue) in rat brains on POD 28. TfR-positive cells were colocalized with NeuN- or CD31- or Olig2-positive cells; however, they were largely observed in the WMI group than in the sham group. White arrows indicate TfR+NeuN+ cells in b5, TfR+CD31+ cells in c5 and d5, and TfR+Olig2+ cells in e5 and f5. Scale bar, 50 μm.

Fig 3: Representative images of immunofluorescence staining for FPN1 (green), NeuN (neuronal marker; red)/CD31 (vascular marker; red)/Olig2 (oligodendrocyte marker; red), and DAPI (nuclear marker; blue) in rat brains on POD 28. FPN1-positive cells were colocalized with NeuN- or CD31- or Olig2-positive cells; however, they were largely observed in the WMI group than in the sham group. White arrows indicate FPN1+NeuN+ cells in a5 and b5, FPN1+CD31+ cells in c5 and d5, and FPN1+Olig2+ cells in e5 and f5. Scale bar, 50 μm.

Fig 4: Representative images of immunofluorescence staining for ferritin (green), NeuN (neuronal marker; red)/CD31 (vascular marker; red)/Olig2 (oligodendrocyte marker; red), and DAPI (nuclear marker; blue) in rat brains on POD 28. Ferritin-positive cells were colocalized with CD31- or Olig2-positive cells; however, they were largely observed in the WMI group than in the sham group. White arrows indicate ferritin+CD31+ cells in c5 and d5 and ferritin+Olig2+ cells in f5. Scale bar, 50 μm.

Fig 5: Chaetocin treatment disrupts GSCs and synergizes with TMZ.(A) qPCR analysis of SUV39H1 and OLIG2 expression in GSCs treated with 0 nM or 50 nM chaetocin. (B and C) Representative images (B) and quantification (C) of tumorsphere formation in GSCs after 48 hours of chaetocin treatment. Unpaired, 2-tailed t test. Scale bars: 100 μm. (D and E) 3D synergy score plots showing the synergistic effect of chaetocin and TMZ on killing GSC3565 (D) and GSC1914 (E). The highest single agent (HSA) synergy score assesses the combination’s efficacy relative to the most effective single agent involved in the combination. An HSA score of greater than 10 is considered indicative of synergy. Data represent mean ± SD. *P < 0.05, **P < 0.01.