cck8 solution  (Boster Bio)

Journal: Experimental Hematology & Oncology

Article Title: The lncRNA PARylator promotes PARP1 activation and resistance to DNA‑damaging therapy in esophageal squamous cell carcinoma

doi: 10.1186/s40164-025-00739-z

Figure Lengend Snippet: PARylator promotes ESCC cell survival and proliferation. a and b SiRNA knockdown of PARylator (a) reduced ESCC cell viability (b) as measured using qPCR (a) and CCK-8 assays (b), respectively. Co-transfection of the PARylator siRNA with an siRNA-resistant PARylator expression plasmid rescued both PARylator expression (a) and cell viability (b). Data shown are mean ± s.d.; n = 3 independent experiments, one-way ANOVA followed by Tukey’s multiple comparison test. c and d Representative photographs (c) and quantification (d) of clonogenic assays in ESCC cells with or without siRNA knockdown of PARylator. Data shown are representatives (c) or mean ± s.d. (d); n = 3 independent experiments, one-way ANOVA followed by Tukey’s multiple comparison test. Scale bar, 1 cm. e SiRNA knockdown of PARylator induced apoptosis in ESCC cells, as measured using flowcytometry analysis of Annexin V/propidium iodide staining, Co-transfection of the PARylator siRNA with an siRNA-resistant PARylator expression plasmid rescued cell apoptosis. Data shown are mean ± s.d.; n = 3 independent experiments, one-way ANOVA followed by Tukey’s multiple comparison test. f SiRNA knockdown of PARylator caused S phase arrest in TE13 cells, as determined by flowcytometry analysis of propidium iodide staining. Data shown are mean ± s.d.; n = 3 independent experiments, one-way ANOVA followed by Tukey’s multiple comparison test. g SiRNA knockdown of PARylator induced caspase-3 activation (cleavage), as measured using Western blotting with an anti-cleaved caspase-3 Ab, Data shown are representatives; n = 3 independent experiments. h SiRNA knockdown of PARylator caused upregulation of PUMA and NOXA, as measured in ESCC cells, as determined by Western blotting. Data shown are representatives; n = 3 independent experiments. i Gene Set Enrichment Analysis (GSEA) of RNA-seq data from TE13 cells showing that knockdown of PARylator using two independent siRNA caused enrichment of the HALLMARK_APOPTOSIS signature. n = 3 experimental repeats. j Induced knockdown of PARylator by treatment with doxycycline (Dox, 200 ng/ml), with knockdown effects attenuated upon Dox withdrawal in TE13.shPARylator and ECA109.PARylator cells. Data shown are mean ± s.d.; n = 3 independent experiments, two-tailed Student’s t -test. k and l Representative photographs (k) and quantification (l) of clonogenic assays in TE13.shPARylator and ECA109.PARylator cells, with or without induced knockdown of PARylator by treatment with doxycycline (Dox, 200 ng/ml). Cessation of Dox treatment led to restoration of PARylator levels and recovery of clonogenic growth. Data shown are representatives (k) or mean ± s.d. (l); n = 3 independent experiments, two-tailed Student’s t -test. Scale bar, 1 cm

Article Snippet: Cell Counting Kit-8 (CCK8) solution (Med Chem Express, #HY-K0301) was added and incubated at 37 °C for 2 h. The absorbance at 450 nm was recorded using a Varioskan LUX microplate reader (ThermoFisher Scientific).

Techniques: Knockdown, CCK-8 Assay, Cotransfection, Expressing, Plasmid Preparation, Comparison, Staining, Activation Assay, Western Blot, RNA Sequencing, Two Tailed Test

Journal: Experimental Hematology & Oncology

Article Title: The lncRNA PARylator promotes PARP1 activation and resistance to DNA‑damaging therapy in esophageal squamous cell carcinoma

doi: 10.1186/s40164-025-00739-z

Figure Lengend Snippet: PARylator knockdown reduces ESCC growth and sensitizes ESCC cells to DNA damage. a and b Representative photographs (a) and growth curves (b) of ECA109.shPARylator xenografts in nu/nu mice ( n = 6 tumors/group), treated as indicated. Data shown are representatives (a) or mean ± s.d. (b); one-way ANOVA followed by Tukey’s multiple comparisons test. c and d Representative microscopic photographs (c) and quantification (d) of Ki67 staining on randomly selected ECA109.shPARylator tumors ( n = 3 tumors) from mice treated as indicated. Data shown are representatives (c) or mean ± s.d. (d); one-way ANOVA followed by Tukey’s multiple comparison test. IRS: immunoreactive score. Scale bar, 20 μm. e PARylator expression in representative ECA109.shPARylator xenografts ( n = 3 tumors) from mice treated as indicated, measured by qPCR. Data shown are mean ± s.d.; one-way ANOVA followed by Tukey’s multiple comparison test. f Cell viability of KYSE450 cells treated with CDDP (0.25 µg/ml, 48 h), cells were subjected to: (i) Control siRNA, (ii) PARylator siRNA, or (iii) PARylator siRNA followed by rescue with an siRNA-resistant PARylator expression plasmid. Data shown are mean ± s.d.; n = 3 independent experiments, one-way ANOVA followed by Tukey’s multiple comparison test. g SiRNA knockdown of PARylator sensitized ESCC cells to IR (4 Gy, single fraction) in TE13 and KYSE450 cells, as measured by CCK-8 assays. Data shown are mean ± s.d.; one-way ANOVA followed by Tukey’s multiple comparison test. h and i Representative microphotographs (h) and quantification (i) of comet tails in TE13 cells treated with or without CDDP (0.25 µg/ml, 48 h), cells were subjected to: (i) Control siRNA, (ii) PARylator siRNA, or (iii) PARylator siRNA followed by rescue with an siRNA-resistant PARylator expression plasmid. Data shown are representatives (h) or mean ± s.d. (i); n = 3 independent experiments, one-way ANOVA followed by Tukey’s multiple comparison test. Scale bar, 20 μm. j and k Representative microphotographs (j) and quantification (k) of immunofluorescence staining of γH2AX in TE13 cells treated with or without CDDP (0.25 µg/ml, 48 h), cells were subjected to: (i) Control siRNA, (ii) PARylator siRNA, or (iii) PARylator siRNA followed by rescue with an siRNA-resistant PARylator expression plasmid. Data shown are representatives (j) or mean ± s.d. (k); n = 3 independent experiments, one-way ANOVA followed by Tukey’s multiple comparison test. Scale bar, 5 μm

Article Snippet: Cell Counting Kit-8 (CCK8) solution (Med Chem Express, #HY-K0301) was added and incubated at 37 °C for 2 h. The absorbance at 450 nm was recorded using a Varioskan LUX microplate reader (ThermoFisher Scientific).

Techniques: Knockdown, Staining, Comparison, Expressing, Control, Plasmid Preparation, CCK-8 Assay, Immunofluorescence

Journal: Breast Cancer : Targets and Therapy

Article Title: Dexmedetomidine Suppresses Mitochondrial Autophagy and Apoptosis While Promoting Proliferation in Breast Cancer Cells in vitro via PI3K/AKT Signaling

doi: 10.2147/BCTT.S543090

Figure Lengend Snippet: Effects of dexmedetomidine (DEX) at different concentrations (25, 50, 100 ng/mL) on proliferation, viability, autophagy and apoptosis of breast cancer cells. ( A ) CCK-8 was performed to detect the viability of breast cancer cells treated with different concentrations of DEX for 24, 48, 72 h. ( B and C ) Colony formation assay was used to detect the proliferation ability of cancer cells treated with DEX at different concentrations. (40×) ( D and H ) The expression of LC3-II/LC3-I and PINK1/Parkin protein in breast cancer cells treated with DEX at different concentrations was measured by Western blot. ( I and J ) The apoptosis of breast cancer cells treated with DEX at different concentrations was determined by flow cytometry. * P <0.05, ** P <0.01, *** P <0.001 vs Control. n=3 biological replicates.

Article Snippet: The cells were cultivated for 24, 48, and 72h at 37°C with 5% CO 2 , and later cultured with 10 μL of CCK8 solution (HY-K0301, MCE, USA) in the incubator (Forma Steri-Cult, Thermofisher, USA) at 37°C for 2 h. Using an enzyme-labeled device (Varioskan LUX, Thermofisher, USA), the absorbance at 450 nm was monitored, and the cell viability was examined.

Techniques: CCK-8 Assay, Colony Assay, Expressing, Western Blot, Flow Cytometry, Control

Journal: Breast Cancer : Targets and Therapy

Article Title: Dexmedetomidine Suppresses Mitochondrial Autophagy and Apoptosis While Promoting Proliferation in Breast Cancer Cells in vitro via PI3K/AKT Signaling

doi: 10.2147/BCTT.S543090

Figure Lengend Snippet: Effects of different drug treatments on proliferation, cell viability, autophagy and apoptosis of breast cancer cells. ( A ) CCK-8 was used to detect breast cancer cell viability after treatment with different drugs. ( B, C ) Colony formation assays were conducted to examine the proliferative ability of breast cancer cells. (40×) ( D-H ) The expression of LC3-II/LC3-I and PINK1/Parkin protein in breast cancer cells was detected by western blot. ( I, J ) The apoptosis of breast cancer cells was tested by flow cytometry. * P <0.05, ** P <0.01, *** P <0.001 vs. Control; ^ P <0.05, ^^^ P <0.001 vs. DEX. ++ P <0.01, +++ P <0.001 vs. LY294002. n=3 biological replicates.

Article Snippet: The cells were cultivated for 24, 48, and 72h at 37°C with 5% CO 2 , and later cultured with 10 μL of CCK8 solution (HY-K0301, MCE, USA) in the incubator (Forma Steri-Cult, Thermofisher, USA) at 37°C for 2 h. Using an enzyme-labeled device (Varioskan LUX, Thermofisher, USA), the absorbance at 450 nm was monitored, and the cell viability was examined.

Techniques: CCK-8 Assay, Expressing, Western Blot, Flow Cytometry, Control