pgfp v rs vectors 560  (OriGene)

Journal: Nature Communications

Article Title: Eukaryote initiation factor 6 modulates small-cell lung carcinoma plasticity via the integrin-FAK signaling axis

doi: 10.1038/s41467-026-69899-8

Figure Lengend Snippet: a Enrichment of EMT and NE gene signatures following EIF6 knockdown in non-NE cell states. RNA-seq analysis is performed in n = 2 biologically independent samples. b Gene expression profiling of EMT- and NE-associated pathways upon EIF6 knockdown in non-NE cell states. RNA-seq analysis is performed in n = 2 biologically independent samples. c Immunoblot analysis of EMT-associated and NE proteins following EIF6 knockdown across multiple SCLC models. Blots are representative of n = 3 biologically independent experiments. For H69M, the samples derive from the same experiment but different gels for ZEB1, another for ZEB2, another for MYCN, another for YAP1, another for eIF6 and c-Myc, another for ASCL1, NEUROD1 and β-tubulin were processed in parallel; For H82R, the samples derive from the same experiment but different gels for ZEB2, ZEB1, MYCN and c-Myc, another for YAP1 and ASCL1, another for NEUROD1, eIF6 and β-tubulin were processed in parallel. For H209A, the samples derive from the same experiment, but different gels for ZEB2, ZEB1, MYCN and c-Myc, another for YAP1 and ASCL1, another for eIF6, another for NEUROD1 and β-tubulin were processed in parallel. d H69 cells with EIF6 knockdown were treated with HGF (40 ng mL⁻¹) for two weeks. Bright-field images show adherent growth (top). Attached colonies were stained with crystal violet for quantification ( n = 3 biologically independent experiments). Scale bar, 100 μm. Error bars represent s.d. Statistical significance was assessed using a two-sided unpaired Wilcoxon rank-sum test. e Re-expression of shRNA-resistant wild-type EIF6 using pCDH lentivirus restores HGF-induced cell attachment in EIF6 knockdown cells. Top: immunoblot showing EIF6 knockdown and re-expression efficiency. Bottom, quantification of crystal violet–stained colonies ( n = 4 independent experiments). Error bars represent s.d. Statistical significance was assessed using a two-sided unpaired Wilcoxon rank-sum test. f Representative images of H69 cells with EIF6 knockdown and re-expression after HGF treatment (40 ng mL⁻¹) for three weeks. Bright-field images (left) and crystal violet–stained colonies (right) are shown. Scale bar, 20 μm. g Live/dead cell analysis of shNTC and shEIF6 H69M cells treated with or without EP (carboplatin 10 μM and etoposide 2 μM) for 72 h, measured by flow cytometry. Data are mean ± SEM from n = 3 independent experiments. Statistical significance was determined using a two-sided unpaired Wilcoxon rank-sum test. h Chemotherapy sensitivity of subcutaneous xenografts derived from shNTC or shEIF6 H69 cells. Tumour volume changes were assessed after three chemotherapy cycles ( n = 13 tumours per group). Response categories were defined as CR (−100%), PR (−30% to 0%) and PD (>20%). Lowercase letters indicate tumours used for immunohistochemistry in i . Created in BioRender. Shen, S. (2026) https://BioRender.com/5p1ea8h . i Representative immunohistochemistry images of ASCL1, NEUROD1, POU2F3, YAP1 and EIF6 in corresponding tumours from h . Scale bars, 50 μm.

Article Snippet: Lentiviruses were made by co-transfection of the shRNA lentivirus vector and the packaging plasmid psPAX2 (addgene#12260) and pMD2.G (addgene#12259) through calcium phosphate co-precipitation into 293 T cells.

Techniques: Knockdown, RNA Sequencing, Gene Expression, Western Blot, Staining, Expressing, shRNA, Cell Attachment Assay, Cell Analysis, Flow Cytometry, Derivative Assay, Immunohistochemistry

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: LFS_primary cells transduced with shSCRAMBLE vector (Plasmid #1864, Addgene) were used as a negative control.

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