Fig 1: SCD1 and SCD5 are essential for GSC maintenance and tumorigenesis.(A) Heatmap of SCD5, SCD1, and stemness marker expression across glioma cell lines, neurospherecultured GSCs, and primary GSCs (GDS3885 microarray dataset).(B) Spearman correlation analysis between SCD isoforms and stemness markers using the same transcriptomic data.(C) Morphological changes in GSCs following 6-day BMP4 differentiation (20 ng/mL). Scale bar: 100 μm.(D) Heatmap of stemness (NES, OLIG1/2, SOX2) and differentiation (GFAP) marker expression post-BMP4 treatment.(E) Immunoblot analysis of SCD1/SCD5 protein levels after BMP4-induced differentiation.(F) Viability assay of GSCs following 4-day SCD1/SCD5 knockdown (mean ± SD; n=4; *p<0.05, **p<0.001, ***p<0.0001, Student’s t-test). Lower panel: mRNA knockdown efficiency heatmap.(G) ELDA of stem cell frequency in M12/M120 GSCs after SCD1/SCD5 knockdown (14 days).(H-I) Rescue experiments: Viability of (H) SCD1-overexpressing GSCs with SCD5 knockdown and (I) SCD5-overexpressing GSCs with SCD1 knockdown (n=4).(J-K) Drug response assays: (J) SCD1i-resistant GSCs (treated with the SCD1 inhibitor CAY10566, 1 μM) with SCD5 knockdown, and (K) SCD1i-sensitive GSCs overexpressing SCD5 (n = 4).(L) In vivo tumor growth measured by Fluc bioluminescence imaging of control (SCR) vs. SCD5-knockdown GSCs (n=7 mice/group) at indicated time points.(M) Kaplan-Meier survival curves of mice bearing control or SCD5-deficient GSCs (n=7/group).
Fig 2: SCD1 and SCD5 are essential for GSC maintenance and tumorigenesis(A) Heatmap of SCD5, SCD1, and stemness marker expression across glioma cell lines, neurosphere-cultured GSCs, and primary GSCs (GDS3885 microarray dataset).(B) Spearman correlation analysis between SCD isoforms and stemness markers using the same transcriptomics data.(C) Morphological changes in GSCs following 6-day BMP4 differentiation (20 ng/mL). Scale bar, 100 μm.(D) Heatmap of stemness (NES, OLIG1/2, and SOX2) and differentiation (GFAP) marker expression post-BMP4 treatment.(E) Immunoblot analysis of SCD1/SCD5 protein levels after BMP4-induced differentiation.(F) Viability assay of GSCs following 4-day SCD1/SCD5 knockdown (mean ± SD; n = 4; *p < 0.05, **p < 0.001, and ***p < 0.0001; Student’s t test). Lower: mRNA knockdown efficiency heatmap.(G) ELDA of stem cell frequency in M12/M120 GSCs after SCD1/SCD5 knockdown (14 days).(H) Cell viability assay of GSCs and non-stem glioma cells (NSGCs) generated by FBS-induced differentiation of GSCs following 4 days of SCD5 knockdown (n = Lower: heatmap showing mRNA knockdown efficiency and expression of stemness and differentiation markers.(I and J) Drug response assays: (J) SCD1i-resistant GSCs (treated with the SCD1 inhibitor CAY10566, 1 μM) with SCD5 knockdown, and (K) SCD1i-sensitive GSCs overexpressing SCD5 (n = 4).(K) In vivo tumor growth measured by Fluc bioluminescence imaging of control (SCR) versus SCD5-knockdown GSCs (n = 7 mice/group) at the indicated time points.(L) Kaplan-Meier survival curves of mice bearing control or SCD5-deficient GSCs (n = 7/group).
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