cell counting kit 8 cck 8 solution  (MedChemExpress)

Journal: Non-coding RNA Research

Article Title: SNHG5 enhances colorectal cancer metastasis through RNA–protein interaction with GNB2 and activation of canonical Wnt signaling

doi: 10.1016/j.ncrna.2025.12.002

Figure Lengend Snippet: Highly metastatic MC38-derived CRC cells exhibit enhanced proliferative, migratory, and invasive properties with mesenchymal characteristics . (A) CCK-8 assay demonstrated significantly higher proliferative capacity in the highly metastatic MC38-F3 subline compared to the parental low-metastatic MC38-F0 cells over a 96-h time course. (B) Representative colony formation images and quantification revealed increased clonogenicity in the MC38-F3 subline. (C) Wound healing assays indicated enhanced migratory ability in MC38-F3 cells at 48 h post-scratch. (D) Transwell migration and Matrigel-coated invasion assays showed that MC38-F3 cells exhibited significantly increased motility and invasiveness. Scale bars = 100 μm. (E) Western blot analysis revealed downregulation of the epithelial marker E-cadherin and upregulation of mesenchymal markers N-cadherin, Vimentin, and Slug in MC38-F3 cells. (F) RT-qPCR analysis confirmed significant upregulation of EMT-associated transcription factors (Snail, Slug, Twist1, ZEB1, ZEB2) in MC38-F3 cells. (G) Immunofluorescence staining corroborated the EMT phenotype, showing reduced E-cadherin and increased Vimentin expression in MC38-F3 cells. Nuclei were counterstained with DAPI. Scale bars = 20 μm. Data are presented as mean ± SEM from at least three independent experiments. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001 (Student's t -test).

Article Snippet: Cell proliferation was assessed using the Cell Counting Kit-8 (CCK-8; HY-K0301, MedChemExpress, China) following the manufacturer's instructions.

Techniques: Derivative Assay, CCK-8 Assay, Migration, Western Blot, Marker, Quantitative RT-PCR, Immunofluorescence, Staining, Expressing

Journal: Non-coding RNA Research

Article Title: SNHG5 enhances colorectal cancer metastasis through RNA–protein interaction with GNB2 and activation of canonical Wnt signaling

doi: 10.1016/j.ncrna.2025.12.002

Figure Lengend Snippet: Snhg5 promotes proliferation, inhibits apoptosis, and enhances motility in CRC cells . (A) CCK-8 assays revealed that Snhg5 overexpression significantly promoted cell proliferation in low-metastatic MC38-F0 cells, while knockdown of Snhg5 suppressed proliferation in highly metastatic MC38-F3 cells over 96 h. (B) Colony formation assays showed increased clonogenic potential in Snhg5-overexpressing MC38-F0 cells and reduced colony formation following Snhg5 silencing in MC38-F3 cells. (C) EdU incorporation assays demonstrated elevated DNA synthesis in Snhg5-overexpressing cells and markedly reduced EdU-positive cell fractions upon Snhg5 knockdown. Scale bars = 100 μm. (D) Annexin V–FITC/PI staining followed by flow cytometry showed that Snhg5 overexpression reduced apoptosis, whereas its knockdown significantly increased apoptotic cell populations. (E) Cell cycle analysis indicated that Snhg5 knockdown induced G1 phase arrest and reduced the S-phase fraction, whereas overexpression promoted G1/S transition. (F) Wound healing assays demonstrated enhanced migration in MC38-F0 cells upon Snhg5 overexpression, and impaired migratory capacity in MC38-F3 cells following knockdown. (G) Transwell migration and invasion assays confirmed that Snhg5 facilitated motility and invasiveness in CRC cells. Snhg5 silencing significantly reduced both migratory and invasive capacities. Scale bars = 100 μm. Data are presented as mean ± SEM from at least three independent experiments. Statistical significance was determined using Student's t -test or ANOVA as appropriate. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001.

Article Snippet: Cell proliferation was assessed using the Cell Counting Kit-8 (CCK-8; HY-K0301, MedChemExpress, China) following the manufacturer's instructions.

Techniques: CCK-8 Assay, Over Expression, Knockdown, DNA Synthesis, Staining, Flow Cytometry, Cell Cycle Assay, Migration

Journal: Non-coding RNA Research

Article Title: SNHG5 enhances colorectal cancer metastasis through RNA–protein interaction with GNB2 and activation of canonical Wnt signaling

doi: 10.1016/j.ncrna.2025.12.002

Figure Lengend Snippet: GNB2 overexpression functionally rescues the tumor-suppressive effects of Snhg5 knockdown in highly metastatic CRC cells . (A) CCK-8 assay showed that Snhg5 knockdown significantly suppressed cell viability over a 96-h period in MC38-F3 cells, whereas GNB2 overexpression partially restored proliferation. (B) Colony formation assays demonstrated that the reduction in clonogenicity induced by Snhg5 knockdown was significantly reversed by GNB2 overexpression. (C) EdU incorporation assay revealed that DNA synthesis was markedly decreased following Snhg5 silencing and partially rescued upon GNB2 overexpression. Scale bars = 100 μm. (D) Annexin V–FITC/PI apoptosis assay showed that Snhg5 depletion significantly increased apoptotic cell populations, which were reduced by GNB2 overexpression. (E) Cell cycle analysis revealed that Snhg5 knockdown induced G1 phase arrest and decreased the S-phase fraction, whereas co-expression of GNB2 alleviated these changes. (F) Wound healing assays demonstrated that the impaired migration capacity caused by Snhg5 silencing was significantly restored by GNB2 overexpression at 48 h. (G) Transwell migration and Matrigel-coated invasion assays further confirmed that GNB2 overexpression rescued the reduction in migratory and invasive abilities induced by Snhg5 knockdown. Scale bars = 100 μm. Data are presented as mean ± SEM from at least three independent experiments. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001 (one-way ANOVA with Tukey's post hoc test unless otherwise indicated).

Article Snippet: Cell proliferation was assessed using the Cell Counting Kit-8 (CCK-8; HY-K0301, MedChemExpress, China) following the manufacturer's instructions.

Techniques: Over Expression, Knockdown, CCK-8 Assay, DNA Synthesis, Apoptosis Assay, Cell Cycle Assay, Expressing, Migration

Journal: Genes & Diseases

Article Title: Nuclear and cytoplasmic USP30-AS1 coordinately regulate breast cancer progression through HnRNPF/p21 and EZH2/c-Myc/p21 axes

doi: 10.1016/j.gendis.2025.101684

Figure Lengend Snippet: USP30-AS1 knockdown inhibits breast cancer cell proliferation both in vitro and in vivo . (A) The knockdown efficacy of USP30-AS1 siRNA and shRNA was validated by qRT-PCR analysis. (B) The effect of USP30-AS1 knockdown on MDA-MB-231 cell proliferation was evaluated by CCK-8 assays. (C) The impact of USP30-AS1 knockdown on the proliferative capacity of MDA-MB-231 cells was assessed by colony formation assays. (D) Flow cytometry analysis was performed to determine the effect of USP30-AS1 knockdown on the cell cycle progression of MDA-MB-231 cells. (E) EdU assays were conducted in MDA-MB-231 cells with USP30-AS1 knockdown. (F) A xenograft tumor experiment was conducted using three MDA-MB-231 cell lines: shNC, shUSP30-AS1#1, and shUSP30-AS1#2. (G) Tumor growth and tumor weight were assessed every two days. (H) The actual photo of tumors for each group was collected at the end of the experiment. (I) Hematoxylin and eosin staining of tumor tissues from nude mice. (J) Immunohistochemistry assays were performed to determine the impact of USP30-AS1 knockdown on Ki67 expression within xenograft tumor tissues.

Article Snippet: The cells were seeded into a 96-well plate (2 × 10 3 /well) and cultured for 0, 24, 48, 72, and 96 h. Cells were treated with 10% CCK-8 solution (MedChemExpress, # HY-K0301) and incubated for 2 h. OD 450 was determined using a microplate reader.

Techniques: Knockdown, In Vitro, In Vivo, shRNA, Quantitative RT-PCR, CCK-8 Assay, Flow Cytometry, Staining, Immunohistochemistry, Expressing

Journal: Translational Oncology

Article Title: Ferroptosis induction enhances anti-PD-1 efficacy in NSCLC via HIF-1α/PD-L1 modulation

doi: 10.1016/j.tranon.2026.102685

Figure Lengend Snippet: Ferroptosis inducers cause cell death and affect PD-L1 expression, which can be inhibited by iron chelators. (A, B) A549 and H1299 cells were treated with erastin or RSL3 at different concentration gradients for 24 h, and cell viability was detected using the CCK-8 kit. Subsequently, cells were co-treated with DFO (100 μM), ferrostatin-1 (2 μM), and the aforementioned ferroptosis inducers at different concentrations for another 24 h, with cell viability re-detected by CCK-8.(C) A549 and H1299 cells were treated with erastin (20 μM) and RSL3 (5 μM), respectively, for 0, 3, 6, 12, 18, and 24 h. PD-L1 expression was measured by qPCR.(D) After A549 and H1299 cells were treated with erastin (20 μM) for 24 h, they were further treated with DFO (100 μM) and ferrostatin-1 (2 μM) alone or in combination. PD-L1 expression in cells was then detected. (E) A549 and H1299 cells were treated with erastin (20 μM) for 24 h, and intracellular ROS was detected by flow cytometry. CCK-8, cell counting kit-8; DFO, deferoxamine; qPCR, quantitative polymerase chain reaction; PD-L1, programmed death-ligand 1; ROS, reactive oxygen species. *** p < 0.001.

Article Snippet: Reagents and Antibodies Cell culture media and reagents: Ham's F-12 K, DMEM, RPMI-1640 (Servicebio), BCA protein assay kit, CCK-8 kit, RIPA lysis buffer (Servicebio), Lovastatin, Fluvastatin, RSL3, Erastin (MCE), deferoxamine mesylate, and Ferrostatin-1 (MCE).

Techniques: Expressing, Concentration Assay, CCK-8 Assay, Flow Cytometry, Cell Counting, Real-time Polymerase Chain Reaction