cck 8 reagent  (Keygen Biotech)

Journal: Genes & Diseases

Article Title: Identification of PLXNC1 as a novel biomarker for consensus molecular subtype 4 in colorectal cancer

doi: 10.1016/j.gendis.2025.101974

Figure Lengend Snippet: PLXNC1 promotes the proliferation, migration, and invasion of colorectal cancer (CRC) cells in vitro. (A) qRT-PCR analysis was conducted to examine the knockdown efficiency of PLXNC1 in the negative control (NC)-siRNA- or PLXNC1-siRNA-transfected CRC cells. NC-siRNA was used as a negative control. GAPDH was a normalization control. (B) CCK-8 assay was performed to assay the viability of control and PLXNC1-silenced CRC cells. (C) After 48 h of transfection with NC-siRNA or PLXNC1-siRNA, the migration of CRC cells in Transwell chambers was determined. (D) After 48 h of transfection with NC-siRNA or PLXNC1-siRNA, CRC cells invading the Matrigel in Transwell chambers were performed. Note: Scale bars = 200 μm. (E) Representative images of EdU-stained (red) cells and the quantification of cell proliferation ratio normalized to Hoechst-stained (blue) cells. Scale bars = 75 μm. (F) qRT-PCR data of the PLXNC1 expression in LoVo cells following transfection with PLXNC1 plasmids or controls. GAPDH was a normalization control. (G) CCK-8 assay results show the viability of LoVo cells following transfection with PLXNC1 plasmids or controls. (H, I) Transwell assay was employed to count the number of migrated (H) and invaded (I) LoVo cells following transfection with PLXNC1 plasmids or controls. ∗∗∗ p < 0.001; ∗∗ p < 0.01, and ∗ p < 0.05.

Article Snippet: CCK-8 reagent (10 μL, DOJINDO, Kumamoto, Japan) was added to each well at 0, 24, 48, and 72 h after transfection and incubated at 37 °C for 4 h. Absorbance at 450 nm was measured using a microplate reader (PerkinElmer EnVision, Massachusetts, USA).

Techniques: Migration, In Vitro, Quantitative RT-PCR, Knockdown, Negative Control, Transfection, Control, CCK-8 Assay, Staining, Expressing, Transwell Assay

Journal: Materials Today Bio

Article Title: Hepatic cavernous hemangioma decellularized extracellular matrix/GelMA composite hydrogel promotes angiogenesis via the ITGA9–FAK–ERK1/2 axis

doi: 10.1016/j.mtbio.2026.102976

Figure Lengend Snippet: Biological responses of HUVECs to HCH dECM. (A) Live/dead staining of HUVECs cultured on HCH dECM/GelMA composite hydrogels (HCH gel) with varying dECM concentrations (scale bars = 200 μm). (B) Quantification of cell viability from graph A. (C) CCK-8 assay assessing HUVEC proliferation in response to HCH gel. (D) Scratch wound assay demonstrating HUVEC migration (scale bars = 200 μm). (E) Transwell migration assay further evaluating HUVEC migration (scale bars = 200 μm). (F) Tube formation assay assessing the angiogenic potential of HUVECs (scale bars = 200 μm). (G) Wound closure rate quantified from graph D. (H) Number of migrating cells quantified from graph E. (I) Total tube length of vessel-like structures quantified from graph F. (n = 5, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001).

Article Snippet: After 1, 2, and 3 days, the culture medium was replaced with 100 μL of fresh medium containing 10% (v/v) CCK-8 reagent (Solarbio, China) and incubated for 2 h at 37 °C.

Techniques: Staining, Cell Culture, CCK-8 Assay, Scratch Wound Assay Assay, Migration, Transwell Migration Assay, Tube Formation Assay

Journal: Materials Today Bio

Article Title: Hepatic cavernous hemangioma decellularized extracellular matrix/GelMA composite hydrogel promotes angiogenesis via the ITGA9–FAK–ERK1/2 axis

doi: 10.1016/j.mtbio.2026.102976

Figure Lengend Snippet: Mechanism underlying the regulation of HUVEC behavior by the HCH gel. (A) Western blot analysis of ITGA9 expression and the phosphorylation status of FAK and ERK1/2. (B) Quantification of Western blot results (n = 3). (C) CCK-8 assay detecting the effect of ITGA9 knockdown (KD) on the proliferation of HUVECs cultured on the HCH gel (n = 5). (D) Transwell migration assay evaluating the effect of ITGA9 KD on HUVEC migration (scale bars = 200 μm). (E) Quantification of the number of migrating cells from graph D (n = 5). (F) Tube formation assay assessing the effect of ITGA9 KD on the angiogenic capacity of HUVECs (incubation for 12 h, scale bars = 200 μm). (G) Quantification of the total tube length from graph F (n = 5). (∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001).

Article Snippet: After 1, 2, and 3 days, the culture medium was replaced with 100 μL of fresh medium containing 10% (v/v) CCK-8 reagent (Solarbio, China) and incubated for 2 h at 37 °C.

Techniques: Western Blot, Expressing, Phospho-proteomics, CCK-8 Assay, Knockdown, Cell Culture, Transwell Migration Assay, Migration, Tube Formation Assay, Incubation

Journal: IBRO Neuroscience Reports

Article Title: TNKS1 mediates the PTEN-PI3K/AKT pathway to regulate glycolysis and proliferation in gliomas

doi: 10.1016/j.ibneur.2026.01.007

Figure Lengend Snippet: TNKS1 can activate the PI3K/AKT pathway by inhibiting the expression of PTEN in glioma cells.Western blotting was used to detect the expression of PTEN in U87 and U251 cell lines (A); the CCK-8 assay was used to measure the viability of U87 and U251 glioma cells treated with different concentrations of LY294002 and 740 Y-P (B); Western blotting was also used to detect the expression of PI3K, p-AKT/AKT in U87 and U251 cell lines (C).*P < 0.05 vs si-NC，@P < 0.05 vs si-NC+Agonist，#P < 0.05 vs si-TNKS1，&P < 0.05 vs OE-TNKS1.

Article Snippet: Subsequently, 10 μL of CCK-8 reagent (Batch no. KGA317, KeyGen Biotech, Nanjing, China) was added to each well.

Techniques: Expressing, Western Blot, CCK-8 Assay

Journal: Translational Oncology

Article Title: UCHL1 promotes temozolomide resistance in glioblastoma by inhibiting the ubiquitination-mediated degradation of keratin 8

doi: 10.1016/j.tranon.2026.102728

Figure Lengend Snippet: UCHL1 is positively associated with TMZ resistance in glioblastoma cells. (A) Cell viability curves for the U87MG, T98G, and U251 cell lines following TMZ treatment were evaluated using the CCK-8 assay. The IC50 values were determined through nonlinear regression analysis (curve fitting), with three replicates ( n = 3). (B) The IC50 values for TMZ-treated U251 cells were assessed after exposure to 100 nM MG132 (a proteasome inhibitor) or 5 μM CQ (an autophagy/lysosome inhibitor) ( n = 3). (C) Differential genes from the datasets GSE193957 , GSE211272 , GSE229600 , and GSE113510 were intersected with members of the ubiquitin proteasome family. Western blotting analysis (D) and RT-qPCR (E) were conducted to assess UCHL1 levels in the U87MG, T98G, and U251 cell lines ( n = 3). * p < 0.05, *** p < 0.001.

Article Snippet: After 48 h of incubation, 10% (v/v) Cell counting kit-8 (CCK-8) reagent (Dojindo, Kumamoto, Japan) was added to each well.

Techniques: CCK-8 Assay, Ubiquitin Proteomics, Western Blot, Quantitative RT-PCR

Journal: Brain, Behavior, & Immunity - Health

Article Title: Physical activity-associated extracellular vesicles inhibit inflammagen-induced microglial activation

doi: 10.1016/j.bbih.2026.101200

Figure Lengend Snippet: Plasma derived from TMPA rats inhibits the LPS-induced activation of NF-κB in BV2 microglial cells. (A) Timeline for experiments identifying the optimal dosage of LPS treatment for BV2 cell. (B) Representative Western blot images showing phosphorylated levels of molecules involved in MAPK and NF-κB signaling in BV2 cells treated with various LPS doses at indicated time points. (C) Quantitative results of phosphorylated level of p65 in BV2 cells treated with various LPS doses at indicated time points. (D) Quantitative results of phosphorylated level of JNK in BV2 cells treated with various LPS doses at indicated time points. (E) Timeline for experiments evaluating the effects of plasma from SED and TMPA rats on p65 and JNK phosphorylation in BV2 cells treated with 100 ng/mL of LPS. (F) Quantitative results of cell viability using CCK-8 assay. (G) Representative Western blot images showing phosphorylated levels of p65 and JNK in BV2 cells treated with plasma from SED or TMPA rats followed by 100 ng/mL of LPS at indicated time points. (H) Quantitative results of phosphorylated level of p65 in BV2 cells treated with plasma from SED or TMPA rats followed by 100 ng/mL of LPS at indicated time points. (I) Quantitative results of phosphorylated level of JNK in BV2 cells treated with plasma from SED or TMPA rats followed by 100 ng/mL of LPS at indicated time points. Data in panels (C) and (D) were expressed as the median with interquartile range and analyzed with Kruskal-Wallis test followed by Dunn's multiple comparisons test. Data in panels (F) , (G) and (H) were expressed as mean ± standard deviation and analyzed with ordinary two-way ANOVA followed by Tukey's multiple comparisons test. Statistical significances from post hoc multiple comparisons were indicated by asterisks: * p < 0.05, ** p < 0.01, *** p < 0.001. Asterisks in panels (C) and (D) indicate significant differences compared to the 0 ng/mL group. n = 4 cultures/group in assays shown in panels (B – D) and n = 9 cultures/group in assays shown in panels (F – I) .

Article Snippet: One hour before the end of treatment, 22 μL of CCK-8 reagent (Cat#: C0005, TargetMol, Boston, MA, USA) was added to each well and incubated for 1 h at 37 °C.

Techniques: Clinical Proteomics, Derivative Assay, Activation Assay, Western Blot, Phospho-proteomics, CCK-8 Assay, Standard Deviation

Journal: Brain, Behavior, & Immunity - Health

Article Title: Physical activity-associated extracellular vesicles inhibit inflammagen-induced microglial activation

doi: 10.1016/j.bbih.2026.101200

Figure Lengend Snippet: Circulating EVs derived from TMPA rats inhibits the LPS-induced activation of NF-κB in BV2 microglial cells. (A) Representative fluorescence images from the EV uptake assay in BV2 cells. The images in the rightmost column are magnified views of the areas outlined by red frames in the color-merged images. Scale bar: 50 μm. (B) Quantitative results of EV uptake assay in BV2 cells. (C) Timeline for experiments evaluating the effects of circulating EVs from SED and TMPA rats on p65 and JNK phosphorylation in BV2 cells treated with 100 ng/mL of LPS. (D) Quantitative results of cell viability using CCK-8 assay. (E) Representative Western blot images showing phosphorylated levels of p65 and JNK in BV2 cells treated with circulating EVs from SED or TMPA rats followed by 100 ng/mL of LPS at indicated time points. (F) Quantitative results of phosphorylated level of p65 in BV2 cells treated with circulating EVs from SED or TMPA rats followed by 100 ng/mL of LPS at indicated time points. (G) Quantitative results of phosphorylated level of JNK in BV2 cells treated with circulating EVs from SED or TMPA rats followed by 100 ng/mL of LPS at indicated time points. Data in panel (B) was expressed as the median with 95% confidence interval. Data in panels (D) , (F) and (G) were expressed as mean ± standard deviation and analyzed with ordinary two-way ANOVA followed by Tukey's multiple comparisons test. Statistical significances from post hoc multiple comparisons were indicated by asterisks: *** p < 0.001. n = 3 cultures/group in assays shown in panels (A) and (B) and n = 9 cultures/group in assays shown in panels (D – G) . (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: One hour before the end of treatment, 22 μL of CCK-8 reagent (Cat#: C0005, TargetMol, Boston, MA, USA) was added to each well and incubated for 1 h at 37 °C.

Techniques: Derivative Assay, Activation Assay, Fluorescence, Phospho-proteomics, CCK-8 Assay, Western Blot, Standard Deviation

Journal: Cancer Biology & Therapy

Article Title: Parthenolide inhibits the progression of intrahepatic cholangiocarcinoma by promoting ferroptosis through inhibiting UBD

doi: 10.1080/15384047.2026.2664327

Figure Lengend Snippet: Inhibition of UBD and Parthenolide treatment synergistically promotes ferroptosis in ICC. (A) qRT-PCR analysis of UBD mRNA expression levels in cells transfected with si-UBD constructs and UBD overexpression plasmid. (B) Western blot analysis of UBD protein expression levels in cells transfected with si-UBD constructs and UBD overexpression plasmid. (C) CCK-8 assay results showing cell viability (OD value at 450 nm) in cells with UBD knockdown (Si-1-UBD), overexpression (OE-UBD), and cells treated with the Par. (D) CCK-8 assay results showing cell viability in cells with UBD knockdown, treated with Par alone, or co-treated with Fer-1. (E–G) Quantification of intracellular relative iron level (E), MDA level (F), and SOD activity (G) in Si-Ctrl, Si-1-UBD, Si-Ctrl + Par-5, and Si-1-UBD + Par-5 groups. Note: Par: Parthenolide; * p < 0.05, ** p < 0.01, *** p < 0.005, ns: not significant.

Article Snippet: The CCK-8 reagent kit (#CK04) was purchased from Dojindo (Kumamoto Prefecture, Japan).

Techniques: Inhibition, Quantitative RT-PCR, Expressing, Transfection, Construct, Over Expression, Plasmid Preparation, Western Blot, CCK-8 Assay, Knockdown, Activity Assay