cck 8 reagent  (MedChemExpress)

Journal: Biochemistry and Biophysics Reports

Article Title: Bifendate inhibits cell PARthanatos by activating the MEK/ERK pathway

doi: 10.1016/j.bbrep.2026.102454

Figure Lengend Snippet: DDB suppresses MNNG-induced PARthanatos. (A) Chemical structure of DDB (ChemSpider). (B) Representative images of HeLa cells pre-treated with vehicle or the indicated concentrations of DDB for 24 h, exposed to MNNG (60 μM, 15 min), further incubated with DDB for 12 h, and finally stained with Sytox-Green-containing medium for 30 min. (C) HeLa cells were pre-treated with graded concentrations of DDB for 24 h, challenged with MNNG (60 μM, 15 min), and cultured for an additional 24 h in the presence of DDB. Cell viability was then measured with the CCK-8 assay. (D–E) Scale bar: 50 μm. The values are expressed as the means ± SDs (n = 3). Statistical analysis was performed using one-way ANOVA followed by Tukey's test. ****p < 0.0001.

Article Snippet: After the indicated treatments, 10 μL CCK-8 reagent (TargetMol, #C0005) was added to each well, and incubated for 2 h at 37 °C.

Techniques: Incubation, Staining, Cell Culture, CCK-8 Assay

Journal: Biochemistry and Biophysics Reports

Article Title: Bifendate inhibits cell PARthanatos by activating the MEK/ERK pathway

doi: 10.1016/j.bbrep.2026.102454

Figure Lengend Snippet: Screening of three DDB analogues. (A–C) Chemical structures of Bicyclol, Schisandrin A and Schisandrin B; all structures were obtained from ChemSpider. (D–E) Representative micrographs (D) and CCK-8 viability results (E) of HeLa cells treated with Schisandrin A or Schisandrin B for 24 h, exposed to MNNG (60 μM, 15 min) and cultured for a further 24 h. Scale bar: 100 μm. (F–G) Representative images (F) and CCK-8 absorbance (G) of HeLa cells pre-treated with graded concentrations of Bicyclol (0–100 μM), challenged with MNNG (60 μM, 15 min) and cultured for 24 h. Scale bar: 25 μm. The values are expressed as the means ± SDs (n = 3). Statistical analysis was performed using one-way ANOVA followed by Tukey's test. ****p < 0.0001.

Article Snippet: After the indicated treatments, 10 μL CCK-8 reagent (TargetMol, #C0005) was added to each well, and incubated for 2 h at 37 °C.

Techniques: Analogues, CCK-8 Assay, Cell Culture

Journal: Biochemistry and Biophysics Reports

Article Title: Bifendate inhibits cell PARthanatos by activating the MEK/ERK pathway

doi: 10.1016/j.bbrep.2026.102454

Figure Lengend Snippet: DDB markedly suppresses PARthanatos in a neuronal cell model. (A–B) SH-SY5Y cells were pre-treated for 24 h with the death inhibitors olaparib (30 μM), Z-VAD (20 μM), necrostatin-1 (20 μM), 3-MA (2 mM) or DDB (25 μM), exposed to MNNG (60 μM, 15 min) and cultured for a further 24 h in the continuous presence of the drugs. Representative micrographs (A) and CCK-8 viability (B) are shown. (C–D) SH-SY5Y cells were exposed to MNNG (60 μM, 15 min) and cultured for 6 h. Representative confocal images (C) and line-scan fluorescence intensity profiles (D) show the effect of DDB on MNNG-induced AIF translocation. Profiles generated with ImageJ. (E) Immunoblot analysis of ERK1 and phospho-ERK in SH-SY5Y cells harvested 0.5, 4 and 6 h after MNNG exposure. (F–G) Representative micrographs (F) and CCK-8 viability (G) after 24 h treatment with DDB (25 μM) alone or in combination with the ERK inhibitor Ravoxertinib (1 μM) in the presence or absence of MNNG (60 μM, 15 min). (H) ERK1 expression levels in SH-SY5Y and HeLa cells. Scale bar: 100 μm. The values are expressed as the means ± SDs (n = 3). Statistical analysis was performed using one-way ANOVA followed by Tukey's test. ****p < 0.0001.

Article Snippet: After the indicated treatments, 10 μL CCK-8 reagent (TargetMol, #C0005) was added to each well, and incubated for 2 h at 37 °C.

Techniques: Cell Culture, CCK-8 Assay, Fluorescence, Translocation Assay, Generated, Western Blot, Expressing

Journal: Cancer Communications

Article Title: TIMELESS Promotes LUAD Growth via Suppressing Transferrin-Mediated Ferroptosis and Reprograms the Tumor Microenvironment against Anti-PD-1 Immunotherapy

doi: 10.34133/cancomm.0009

Figure Lengend Snippet: Dysregulated RNA-binding proteins in LUAD: from multicohort screening to functional validation. (A to D) Volcano plots showing the DE RBPs in the following 4 LUAD cohorts: The Cancer Genome Atlas Lung Adenocarcinoma (TCGA-LUAD) dataset ( n = 568; A), GSE32863 ( n = 116; B), GSE40419 ( n = 164; C), and GSE75037 ( n = 166; D). The significantly DE RBPs ( P adj < 0.05, |log 2 FC| > 1) are shown in blue, and the nonsignificantly DE RBPs ( P adj > 0.05, |log 2 FC| > 1) are shown in dark gray. The other non-RBP genes are shown in light gray. (E and F) Venn diagram showing the intersection of up-regulated (E) and down-regulated (F) DE RBPs across TCGA-LUAD and the 3 GEO datasets. (G) The volcano plot displayed the hazard ratios for patients stratified by the median expression level of RBP genes in the TCGA-LUAD cohort (red: up-regulated RBPs in tumors; blue: down-regulated RBPs in tumors). (H) Heatmap of significantly dysregulated RBPs identified in (E) and (F). The top 10 highest-expressed RBPs in tumors are highlighted (ranked by average tumor z score). (I) Expression levels of the top 10 LUAD-up-regulated RBPs in LUAD cell lines (A549 and H1975) with or without siRNA-mediated knockdown ( n = 4 each group). (J) Calcein-AM/PI staining showed the percentage of viable cells in the negative control group and following the knockdown of the RBP gene ( n = 4 each group). (K) CCK-8 assay of cell viability in RBP gene knockdown and control cells at the indicated time points. The statistical analysis was performed using a 2-tailed Student’s t test (I and J) or 2-way ANOVA (K). *P < 0.05, **P < 0.01, ***P < 0.001, data without statistically significant differences are not labeled (J and K). Abbreviations: ALDH18A1, aldehyde dehydrogenase 18 family member A1; BZW2, basic leucine zipper and W2 domains 2; CCK-8, cell counting kit-8; CENPF, centromere protein F; DE, differentially expressed; FAM83A, family with sequence similarity 83 member A; FC, fold change; FDR, false discovery rate; GEO, Gene Expression Omnibus; IF, immunofluorescence; IHC, immunohistochemistry; KIF20A, kinesin family member 20A; LUAD, lung adenocarcinoma; OS, overall survival; PDIA4, protein disulfide isomerase family A member 4; PRC1, protein regulator of cytokinesis 1; RBP, RNA-binding protein; SRPK1, SRSF protein kinase 1; TCGA, The Cancer Genome Atlas; TMA, tissue microarray; TOP2A, DNA topoisomerase II alpha.

Article Snippet: Cell counting kit-8 (CCK-8) reagent (MedChemExpress, Cat. HY-K0301) was added to each well, and the absorbance at 450 nm was measured using a microplate reader (TECAN, Infinite 200 PRO).

Techniques: RNA Binding Assay, Functional Assay, Biomarker Discovery, Expressing, Knockdown, Staining, Negative Control, CCK-8 Assay, Control, Labeling, Cell Counting, Sequencing, Gene Expression, Immunofluorescence, Immunohistochemistry, Microarray