Journal: Neoplasia (New York, N.Y.)

Article Title: A MYC family switch: L-MYC drives and maintains neuroendocrine lineage programs in prostate cancer

doi: 10.1016/j.neo.2026.101307

Figure Lengend Snippet: MYCL reduces proliferation while promoting migration and cytoskeletal remodeling and decreasing cell adhesion in prostate cancer cells (A) Proliferation assay. Real-time Incucyte analysis showing cell proliferation of C4-2B control (Ctrl) and C4-2B-MYCL cells measured as confluence (%) over time. Quantification using area under the curve (AUC) demonstrates reduced proliferative capacity following MYCL overexpression. (B) Proliferation assay. Real-time Incucyte analysis showing cell proliferation of PC3 control (Ctrl) and PC3-MYCL cells measured as confluence (%) over time. Quantification using area under the curve (AUC) demonstrates reduced proliferative capacity following MYCL overexpression. (C) Left: Cell cycle analysis of C4-2B and PC3 cells following MYCL overexpression. In PC3 cells, MYCL reduces the G1 population and increases S-phase, indicating altered cell cycle progression, whereas changes in C4-2B cells are minimal. Right: Apoptosis analysis by Annexin V/7-AAD staining. MYCL overexpression in PC3 cells increases the early apoptotic population, with little or no significant change in C4-2B cells. (D) Cell adhesion assay. Cell adhesion was assessed by crystal violet staining and quantified by measuring absorbance at 595 nm at 18, 24, 48, and 72 h following seeding. MYCL-overexpressing C4-2B cells exhibited significantly decreased adhesion compared with control cells. Representative phase-contrast images acquired 48 h after seeding show reduced cell attachment and increased cell clustering in MYCL-expressing cells. (E) Molecular validation. RT-qPCR analysis of adhesion-related genes (ITGB1, ITGAV) in control (Ctrl) and MYCL-overexpressing C4-2B, LNCaP and PC3 cells. (F) Migration assay. Wound-healing analysis measuring relative wound density (%) over time demonstrates moderately enhanced migratory capacity in PC3-MYCL cells. AUC quantification confirms increased migration upon MYCL expression. (G) Cytoskeletal transcriptional programs. Heatmap showing differential expression of genes meeting thresholds of |log₂FC| ≥ 0.5 and adjusted p-value (FDR) < 0.05. Differentially regulated genes are associated with Ephrin-EPH signaling, Rho-Rac signaling, cytoskeletal organization, cell-cell junctions, extracellular matrix (ECM) interactions, and epithelial-mesenchymal transition (EMT) regulators, indicating MYCL-driven cytoskeletal remodeling signatures.

Article Snippet: For live-cell proliferation analysis, cells were seeded in 96-well plates and monitored using the IncuCyte® Live-Cell Analysis System (Sartorius).

Techniques: Migration, Proliferation Assay, Control, Over Expression, Cell Cycle Assay, Staining, Cell Adhesion Assay, Cell Attachment Assay, Expressing, Biomarker Discovery, Quantitative RT-PCR, Quantitative Proteomics

Journal: Neoplasia (New York, N.Y.)

Article Title: A MYC family switch: L-MYC drives and maintains neuroendocrine lineage programs in prostate cancer

doi: 10.1016/j.neo.2026.101307

Figure Lengend Snippet: MYCL reduces proliferation while promoting migration and cytoskeletal remodeling and decreasing cell adhesion in prostate cancer cells (A) Proliferation assay. Real-time Incucyte analysis showing cell proliferation of C4-2B control (Ctrl) and C4-2B-MYCL cells measured as confluence (%) over time. Quantification using area under the curve (AUC) demonstrates reduced proliferative capacity following MYCL overexpression. (B) Proliferation assay. Real-time Incucyte analysis showing cell proliferation of PC3 control (Ctrl) and PC3-MYCL cells measured as confluence (%) over time. Quantification using area under the curve (AUC) demonstrates reduced proliferative capacity following MYCL overexpression. (C) Left: Cell cycle analysis of C4-2B and PC3 cells following MYCL overexpression. In PC3 cells, MYCL reduces the G1 population and increases S-phase, indicating altered cell cycle progression, whereas changes in C4-2B cells are minimal. Right: Apoptosis analysis by Annexin V/7-AAD staining. MYCL overexpression in PC3 cells increases the early apoptotic population, with little or no significant change in C4-2B cells. (D) Cell adhesion assay. Cell adhesion was assessed by crystal violet staining and quantified by measuring absorbance at 595 nm at 18, 24, 48, and 72 h following seeding. MYCL-overexpressing C4-2B cells exhibited significantly decreased adhesion compared with control cells. Representative phase-contrast images acquired 48 h after seeding show reduced cell attachment and increased cell clustering in MYCL-expressing cells. (E) Molecular validation. RT-qPCR analysis of adhesion-related genes (ITGB1, ITGAV) in control (Ctrl) and MYCL-overexpressing C4-2B, LNCaP and PC3 cells. (F) Migration assay. Wound-healing analysis measuring relative wound density (%) over time demonstrates moderately enhanced migratory capacity in PC3-MYCL cells. AUC quantification confirms increased migration upon MYCL expression. (G) Cytoskeletal transcriptional programs. Heatmap showing differential expression of genes meeting thresholds of |log₂FC| ≥ 0.5 and adjusted p-value (FDR) < 0.05. Differentially regulated genes are associated with Ephrin-EPH signaling, Rho-Rac signaling, cytoskeletal organization, cell-cell junctions, extracellular matrix (ECM) interactions, and epithelial-mesenchymal transition (EMT) regulators, indicating MYCL-driven cytoskeletal remodeling signatures.

Article Snippet: Cell migration was evaluated using a wound-healing assay performed with the IncuCyte® system following the manufacturer’s (Sartorius) protocol.

Techniques: Migration, Proliferation Assay, Control, Over Expression, Cell Cycle Assay, Staining, Cell Adhesion Assay, Cell Attachment Assay, Expressing, Biomarker Discovery, Quantitative RT-PCR, Quantitative Proteomics

Journal: STAR Protocols

Article Title: Protocol for Seahorse 3D Mito Stress assay in patient-derived atypical teratoid rhabdoid tumor CHLA-05-ATRT single neurospheres

doi: 10.1016/j.xpro.2026.104515

Figure Lengend Snippet: Oxygen consumption rate (OCR) in the presence or absence of spheroids (A) Incucyte image of spheroid in the center of the microplate well at the end of the assay and (B) the corresponding OCR kinetic graph showing a response after drug injections as expected. (C) Image of a microplate well with no spheroid detected at the end of the assay and (D) corresponding OCR kinetic graph showing no response after drug injections.

Article Snippet: Incucyte Organoid Analysis Software Module , Sartorius , 9600–0034.

Techniques:

Journal: STAR Protocols

Article Title: Protocol for Seahorse 3D Mito Stress assay in patient-derived atypical teratoid rhabdoid tumor CHLA-05-ATRT single neurospheres

doi: 10.1016/j.xpro.2026.104515

Figure Lengend Snippet: Effect of coating timing on spheroid retention in microplate wells (A) Incucyte image of a poly-lysine coated microplate with spheroids transferred on the same day as the assay, taken before the assay. (B) Image of the same plate taken after the assay, showing that the spheroids were displaced and/or destroyed during the assay. (C) Image of a poly-lysine coated microplate with spheroids transferred the day before the assay, taken before the assay and (D) after the assay, showing that the spheroids were intact.

Article Snippet: Incucyte Organoid Analysis Software Module , Sartorius , 9600–0034.

Techniques:

Journal: STAR Protocols

Article Title: Protocol for Seahorse 3D Mito Stress assay in patient-derived atypical teratoid rhabdoid tumor CHLA-05-ATRT single neurospheres

doi: 10.1016/j.xpro.2026.104515

Figure Lengend Snippet: Oxygen consumption rate (OCR) in the presence or absence of spheroids (A) Incucyte image of spheroid in the center of the microplate well at the end of the assay and (B) the corresponding OCR kinetic graph showing a response after drug injections as expected. (C) Image of a microplate well with no spheroid detected at the end of the assay and (D) corresponding OCR kinetic graph showing no response after drug injections.

Article Snippet: Incucyte S3 Live-Cell analysis system , Sartorius , 4647.

Techniques:

Journal: STAR Protocols

Article Title: Protocol for Seahorse 3D Mito Stress assay in patient-derived atypical teratoid rhabdoid tumor CHLA-05-ATRT single neurospheres

doi: 10.1016/j.xpro.2026.104515

Figure Lengend Snippet: Effect of coating timing on spheroid retention in microplate wells (A) Incucyte image of a poly-lysine coated microplate with spheroids transferred on the same day as the assay, taken before the assay. (B) Image of the same plate taken after the assay, showing that the spheroids were displaced and/or destroyed during the assay. (C) Image of a poly-lysine coated microplate with spheroids transferred the day before the assay, taken before the assay and (D) after the assay, showing that the spheroids were intact.

Article Snippet: Incucyte S3 Live-Cell analysis system , Sartorius , 4647.

Techniques: