sc 5285  (Santa Cruz Biotechnology)

Journal: iScience

Article Title: Preclinical drug screen identifies WEE1 inhibitor and vinca alkaloid as a combination treatment concept for Li-Fraumeni syndrome medulloblastoma

doi: 10.1016/j.isci.2025.114564

Figure Lengend Snippet: Drug combination screening and drug hits validation in in vitro TP53 mut brain cancer cell models (A) Average synergy scores estimated for combinations of cell cycle checkpoint kinase inhibitors and micronucleus-associated compounds across TP53 mut brain cancer cell lines; black points indicate the average value across all calculated synergy scores (ATM/ATRi, ATM/ATR inhibitors; CHK1/2i, CHEK1/CHEK2 inhibitors; WEE1i, WEE1 inhibitor; VBL, vinblastine; VCR, vincristine) (see also ). (B) Model representing mechanism of action of adavosertib in TP53 wt and TP53 mut cancer cells exposed to DNA damaging agents (Created with BioRender.com ). (C) Adavosertib (adav) decreases CDK1 phosphorylation levels in SJ-GBM2 and UW228-2 cell lines; Student’s t test was used for statistical analysis, data are represented as mean ± SEM (Δ t , exposure time; ∗ p < 0.05; ∗∗ p < 0.01). (D) WEE1 knockdown (KD) and vincristine (VCR) effect on metabolic activity of in vitro TP53 mut brain cancer cell lines: data are represented as mean ± SEM (siContr, control siRNA; siWEE1, WEE1 siRNA; NS, not significant; ∗ p < 0.05; ∗∗ p < 0.01) (see also C). (E) WEE1 KD and VCR effect on in vitro caspase-3 activity in TP53 mut brain cancer cell lines: data are represented as mean ± SEM (NS, not significant; ∗ p < 0.05; ∗∗ p < 0.01) (see also C). (F) Dose-response curves of WEE1 inhibitors (adavosertib, Debio 0123, and ZN-c3) in in vitro TP53 mut brain cancer cell lines: data are represented as mean ± SEM. (G) Synergy scores of vincristine and WEE1 inhibitors combination treatment in in vitro TP53 mut brain cancer cell lines: data are represented as mean.

Article Snippet: WEE1 knockdown at protein level was confirmed by Western blot (α-WEE1 antibody [B-11], cat.no. sc-5285, Santa Cruz Biotechnology).

Techniques: Biomarker Discovery, In Vitro, Phospho-proteomics, Knockdown, Activity Assay, Control

Journal: iScience

Article Title: Preclinical drug screen identifies WEE1 inhibitor and vinca alkaloid as a combination treatment concept for Li-Fraumeni syndrome medulloblastoma

doi: 10.1016/j.isci.2025.114564

Figure Lengend Snippet: WEE1 as a therapeutic target in pediatric brain tumor entities (A) WEE1 mRNA expression in MB tumors by methylation group (MB dataset: Northcott [ n = 491], cerebellum dataset: Roth [ n = 9] ). (B) WEE1 mRNA expression in MB subgroups (Cavalli [ n = 763] ). (C) Adavosertib sensitivity score in pediatric brain tumors (Petralia [ n = 218] ). (D) Adavosertib sensitivity score in MB groups (Cavalli [ n = 763] ). (E) Adavosertib sensitivity score in MB tumors by methylation group (Cavalli [ n = 763] ) (see also B). Data are represented as mean ± SEM (Student’s t test significance levels: NS, not significant; ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001).

Article Snippet: WEE1 knockdown at protein level was confirmed by Western blot (α-WEE1 antibody [B-11], cat.no. sc-5285, Santa Cruz Biotechnology).

Techniques: Expressing, Methylation

Journal: iScience

Article Title: Preclinical drug screen identifies WEE1 inhibitor and vinca alkaloid as a combination treatment concept for Li-Fraumeni syndrome medulloblastoma

doi: 10.1016/j.isci.2025.114564

Figure Lengend Snippet: In vivo validation of adavosertib and vincristine (VCR) in LFS SHH-MB models (A) Survival of mice injected with BT084 patient-derived xenograft (PDX) model during treatment with adavosertib and VCR; log rank test was used for statistical analysis. (B) Tumor growth dynamics of BT084 PDX model during treatment with adavosertib and vincristine (VCR): data are represented as mean ± SEM. (C) Phospho (p)-CDK1 in LFS SHH-MB PDX cells (HS231222 and LFS primary) following in vivo treatment with adavosertib and VCR: numbers below the blot represent normalized fold change relative to non-treated control. (D) Tumor growth dynamics of LFS MB PDX models expressing WEE1 shRNA (shWEE1) and control shRNA (shSCRAMBLE). (E) Survival of mice injected with LFS MB PDX models expressing shWEE1 and shSCRAMBLE; log rank test was used for statistical analysis.

Article Snippet: WEE1 knockdown at protein level was confirmed by Western blot (α-WEE1 antibody [B-11], cat.no. sc-5285, Santa Cruz Biotechnology).

Techniques: In Vivo, Biomarker Discovery, Injection, Derivative Assay, Control, Expressing, shRNA