Journal: Scientific Reports

Article Title: Glycyrrhizin ameliorates colorectal cancer progression by regulating NHEJ pathway through inhibiting HMGB1-induced DNA damage response

doi: 10.1038/s41598-024-76155-w

Figure Lengend Snippet: Effect of GL mediated cytotoxicity on human colorectal cancer cells. a Percent cell viability at 24 h post-GL treatment for a range of 100–400 µM was assessed by CCK8 assays of HCT116 and HT29 cells. b CCK8 data at 24 h were used to calculate the IC50 values using the nonlinear regression function of GraphPad Prism. GL concentrations are reported in µM on a log (10) scale, DMSO: 0.2% DMSO (vehicle) negative control. c Cell colony formation was measured with or without GL treatment. d Apoptosis was measured using Annexin V/7-AAD double staining in CRC cells 24 h after GL at 300 µM. e Caspase-3 activity was utilized to examine the apoptosis ability of CRC cells post-GL at 300 µM. Data from the three experiments are presented as the mean ± SD. f The protein levels of Caspase-3 were measured via Western blotting in CRC cells after GL treatment. α-Tubulin was used as a loading control. g The effect of GL at 300 µM on the cell cycle distribution in CRC cells. h The protein levels of CDK2 and cyclin D1 were measured via Western blotting in CRC cells after GL treatment. α-Tubulin was used as a loading control. * P < 0.05, ** P < 0.01.

Article Snippet: The antibodies used for Western blotting analysis in this study were as follows: mouse anti-human HMGB1 (Proteintech, #66525-1-Ig), mouse anti-human cyclin D1 (Proteintech, #60186-1-Ig), mouse anti-human CDK2 (Proteintech, #60312-1-Ig), mouse anti-human caspase-3 (Proteintech, #66470-2-Ig), rabbit anti-human/mouse phospho-Histone H 2 A.x-S139 (Abclonal, #AP0099), rabbit anti-human Ku70 (Proteintech, #10723-1-AP), rabbit anti-human LIG4 (Proteintech, #12695-1-AP) and mouse anti-human/mouse α-Tubulin (Proteintech, #66031-1-Ig).

Techniques: Negative Control, Double Staining, Activity Assay, Western Blot, Control

Journal: Scientific Reports

Article Title: Glycyrrhizin ameliorates colorectal cancer progression by regulating NHEJ pathway through inhibiting HMGB1-induced DNA damage response

doi: 10.1038/s41598-024-76155-w

Figure Lengend Snippet: GL treatment alleviates the proliferation of colorectal cancer cells via HMGB1. a The mRNA expression of HMGB1 was measured by qRT‒PCR in CRC cells. Western blotting was used to detect HMGB1 protein expression levels in CRC cells. α-Tubulin was used as a loading control. b Cell viability was measured using CCK-8 assays in HMGB1-overexpressing CRC cells 24 h after GL at 300 µM. c Colony formation assay of HMGB1-overexpressing CRC cells with or without GL treatment. d Apoptosis was measured using Annexin V/7-AAD double staining in HMGB1-overexpressing CRC cells 24 h after GL at 300 µM. e Caspase-3 activity was utilized to examine the apoptosis ability of CRC cells. f Western blotting was used to detect Caspase 3 protein expression levels in HMGB1-overexpression CRC cells post-GL at 300 µM. α-Tubulin was used as a loading control. g Cell cycle distribution was measured in HMGB1-overexpressing CRC cells 24 h after GL treatment at 300 µM. h Western blotting was used to detect CDK2 and cyclin D1 protein expression levels in HMGB1-overexpression CRC cells post-GL at 300 µM. α-Tubulin was used as a loading control. Data from the three experiments are presented as the mean ± SD. * P < 0.05, ** P < 0.01.

Article Snippet: The antibodies used for Western blotting analysis in this study were as follows: mouse anti-human HMGB1 (Proteintech, #66525-1-Ig), mouse anti-human cyclin D1 (Proteintech, #60186-1-Ig), mouse anti-human CDK2 (Proteintech, #60312-1-Ig), mouse anti-human caspase-3 (Proteintech, #66470-2-Ig), rabbit anti-human/mouse phospho-Histone H 2 A.x-S139 (Abclonal, #AP0099), rabbit anti-human Ku70 (Proteintech, #10723-1-AP), rabbit anti-human LIG4 (Proteintech, #12695-1-AP) and mouse anti-human/mouse α-Tubulin (Proteintech, #66031-1-Ig).

Techniques: Expressing, Western Blot, Control, CCK-8 Assay, Colony Assay, Double Staining, Activity Assay, Over Expression