Journal: iScience

Article Title: Acetylation halts missense mutant p53 aggregation and rescues tumor suppression in non-small cell lung cancers

doi: 10.1016/j.isci.2023.107003

Figure Lengend Snippet: Acetylation reverses p53 missense mutation gain-of-functions and rescues tumor suppressive activity (A) Cell growth assay of H1299 cells expressing the indicated constructs (mean ± SD and determined using one-way analysis of variance, n = 6, ∗p < 0.05, ∗∗p < 0.01). (B) Colony formation assay of H1299 cells (mean ± SD and one-way analysis of variance, n = 3, ∗∗p < 0.01). (C) EdU incorporation assay detecting the DNA replication of H1299 cells (mean ± SD and one-way analysis of variance, ∗p < 0.05, ∗∗p < 0.01). Data were expressed as the percentage of EdU positive cells (Red) to total Hoechst-labeled cells (Blue). Bars, 20 μM. (D) Cell cycle detection by flow cytometry and the statistical results (mean ± SD and one-way analysis of variance, n = 3, ∗p < 0.05). (E) Apoptosis assay of H1299 cells. Cells were treated with Dox (0.5 μM) and Eto (10 μM) for 24h and apoptotic cells were analyzed by flow cytometry (mean ± SD and one-way analysis of variance, n = 3, ∗p < 0.05, ∗∗p < 0.01). (F) Representative images of 3D-spheroid growth assay (left) and statistics (right; mean ± SD and one-way analysis of variance, n = 5, ∗∗p < 0.01). Bars, 500 μM. (G) Growth curve of subcutaneous xenografts derived from H1299 cells. The tumor volume was measured at the indicated times after implantation (mean ± SD and one-way analysis of variance, n = 5, ∗∗p < 0.01). (H) Representative images of subcutaneous xenografts collected at the 22nd day after implantation. The right panel shows the tumor weight statistics (mean ± SD and one-way analysis of variance, n = 5, ∗∗p < 0.05). (I) Representative images of p53 and Ac-p53 IHC in non-cancerous tissues (NCT) and LUADs (tumor) tissues. Scale bars, 200 μm in whole images and 10 μm in local enlarged images. (J) Immunoreactivity scoring (IRS) analysis of p53 and Ac-p53 expression (mean ± SD and determined using Student’s t tests, ∗∗p < 0.01). (K) Diagram showing high (Ac-p53H) and low Ac-p53 (Ac-p53L). (L) Overall survival analysis grouped by p53 positive and negative tissues (Log rank). (M) Overall survival analysis grouped by high p53 and low p53 based on IRS cutoff of 6 (Log rank). (N) Overall survival analysis grouped by high Ac-p53 and low Ac-p53 based on IRS cutoff of 8 (Log rank).

Article Snippet: EdU incorporation assay Kit , KeyGen Biotech , Cat# KGA337-500.

Techniques: Mutagenesis, Activity Assay, Growth Assay, Expressing, Construct, Colony Assay, Labeling, Flow Cytometry, Apoptosis Assay, Derivative Assay

Journal: iScience

Article Title: Acetylation halts missense mutant p53 aggregation and rescues tumor suppression in non-small cell lung cancers

doi: 10.1016/j.isci.2023.107003

Figure Lengend Snippet:

Article Snippet: EdU incorporation assay Kit , KeyGen Biotech , Cat# KGA337-500.

Techniques: Recombinant, Chromatin Immunoprecipitation, Sequencing, Software

Journal: Frontiers in Oncology

Article Title: Iron Regulates the Warburg Effect and Ferroptosis in Colorectal Cancer

doi: 10.3389/fonc.2021.614778

Figure Lengend Snippet: Iron promoted CRC cells proliferation and colony formation in vitro . (A) Viabilities of cells treated with different concentrations of FAC (0–1,000 µM) for 12 h. (B) Viabilities of cells treated with FAC (100 µM) or DFO (100 µM) for 0–48 h (C, D) Immunofluorescence images and percentages of Edu incorporation. (E, F) Colony images and numbers of HCT-116 and HT-29 cells. Cells were incubated in medium containing 100 µM FAC or 100 µM DFO for 14 days. (**p < 0.01; ***p < 0.001; ns, not significant).

Article Snippet: An EdU incorporation kit (Beyotime) was used to measure the cell proliferation ability.

Techniques: In Vitro, Immunofluorescence, Incubation

Journal: Frontiers in Oncology

Article Title: Iron Regulates the Warburg Effect and Ferroptosis in Colorectal Cancer

doi: 10.3389/fonc.2021.614778

Figure Lengend Snippet: Iron-induced cell proliferation was rescued by NRF2 knockdown. (A) Western blots of HCT-116 and HT-29 cells transfected with control shRNA (shCtrl) and NRF2 shRNA (shNRF2). shNRF2#1 was used in following studies for its better knockdown effect. (B) Cell viabilities were assessed by CCK-8 kits. Cells were transfected and then treated with FAC (100 µM) or DFO (100 µM) for 12 h. (C, D) Edu assay of transfected HCT-116 and HT-29 cells. Cells were pre-treated with or without 100 µM FAC for 12 h before Edu assay. (E) Colony formation assay of transfected HCT-116 and HT-29 cells. Cells were incubated in medium containing 100 µM FAC or equivalent PBS for 14 days. (**p < 0.01; ***p < 0.001; ns, not significant).

Article Snippet: An EdU incorporation kit (Beyotime) was used to measure the cell proliferation ability.

Techniques: Western Blot, Transfection, shRNA, CCK-8 Assay, EdU Assay, Colony Assay, Incubation

Journal: International Journal of Molecular Sciences

Article Title: Multifaceted Mechanisms of Cisplatin Resistance in Long-Term Treated Urothelial Carcinoma Cell Lines

doi: 10.3390/ijms19020590

Figure Lengend Snippet: Long-term cisplatin treated cells (LTTs) recover from cisplatin-induced stress. ( a ) Cell viability was measured in treatment-naive parental urothelial carcinoma cell lines (UCCs) and resistant LTTs 72 h after treatment with indicated doses of cisplatin by MTT assay. Maintenance cisplatin concentration of LTTs is shown for comparison; ( b ) Proliferation of parental cells and LTTs treated with either IC 50 or maintenance concentrations of cisplatin for 72 h as measured by EdU incorporation assay; ( c ) Colony formation assay of parental cells and LTTs treated with either IC 50 or maintenance concentrations of cisplatin for 72 h. Cell clones were stained by Giemsa; ( d ) Changes in cell-cycle distribution and amount of apoptotic cells (as sub-G1 fraction) in untreated LTTs and 72, 168, and 240 h after maintenance cisplatin treatment measured by flow cytometry. Untreated parental UCCs served as a control (left panel). Values represent the mean ± standard deviation (SD) of two independent experiments. * p < 0.05.

Article Snippet: Cell proliferation was measured by EdU incorporation assay (baseclick GmbH, Neuried, Germany).

Techniques: MTT Assay, Concentration Assay, Colony Assay, Clone Assay, Staining, Flow Cytometry, Standard Deviation