Lues of 143.39 eight.52, 14.75 0.60 and 11.95 0.60 /ml at exposure times of 24, 48 and 72 h, respectively (Figs. 1EF).TLPE extract induces cell cycle arrest in CCA cell linesIn this study, the cholangiocarcinoma cell lines (KKU-M213B, KKU-100) and nontumorigenic biliary epithelial cells (H69) had been treated with TLPE extract at 31.25, 62.50 and 125.00 /ml for 24 h. KKU-M213B cells have been treated with TLPE extract at 31.25, 62.50 and 125.00 /ml for 24 h but didn’t show accumulation of cells at any cell cycle phase.Samankul et al. (2022), PeerJ, DOI ten.7717/peerj.5/Figure 1 The impact of TLPE extract on the proliferation of CCA cells. KKU-M213B (A, B), KKU-100 (C, D) and H69 (D, E) cells have been treated with TLPE extract of 31.2500 /ml for 24 h and three.750 /ml for 48 and 72 h. Antiproliferative activity was determined working with MTT assay as outlined by manufacturer protocol. Cell viabilities are expressed as percentages in the solvent manage, which was defined as 100 cell viability. The IC50 values are indicated because the signifies S.D. from three independent experiments. Full-size DOI: 10.7717/peerj.14518/fig-Samankul et al. (2022), PeerJ, DOI 10.7717/peerj.6/Figure 2 Histograms display the DNA content distribution of CCA cells. KKU-M213B (A), KKU-100 (C) and H69 (E) cell cycle profiles immediately after treated with TLPE extract. Cells treated with 1 EtOH (v/v), 31.25, 62.50 and 125.00 /ml of TLPE extract had been stained with propidium iodide (PI) and analyzed by flow cytometry. Bar graphs represent the summarized information on cell cycle distribution of KKU-M213B (B), KKU-100 (D) and H69 (F) cells. The percentages of cells are presented as suggests S.D. from two independent experiments performed in duplicate. The asterisk “” indicates a substantial boost when compared with solvent control remedies (p 0.05). Full-size DOI: 10.7717/peerj.14518/fig-However, a significant raise in the cell population inside the SubG1 phase was observed up to 12.three , 17.1 and 19.7 , respectively (Figs. 2AB). KKU-100 cells treated with TLPE extract at 31.25, 62.50 and 125.00 /ml for 24 h induced cell cycle arrest at the G0/G1 phase (Figs. 2CD). The cell populations within the G0/G1 phase have been substantially enhanced up to 82.9 , 84.two and 84.six when treated with TLPE extract at 31.25, 62.50 and 125.00 /ml, respectively. Also, a substantial enhance in the cell population inside the SubG1 phase was observed as much as 1.three , 1.7 and 7.three , respectively (Figs. 2CD). H69 cells treated with TLPE extract at 31.25, 62.50 and 125.00 /ml for 24 h showed no accumulation of cells at any cell cycle phase (Figs. 2EF). Nonetheless, a substantial improve on the SubG1 population in H69 cells was observed as much as 7.4 , eight.five and 9.6 , respectively (Figs. 2EF).Samankul et al. (2022), PeerJ, DOI 10.7717/peerj.7/Figure three Apoptotic death evaluation by flow cytometry.Hemoglobin subunit theta-1/HBQ1 Protein Molecular Weight Representative dot plots show the apoptotic death with the cells treated using the indicated concentrations in KKU-M213B (A), KKU-100 (C) and H69 (E) cells treated with TLPE extract.Protease Inhibitor Cocktail custom synthesis Soon after 24 h-treatment, cells labeled with Annexin V-FITC and Propidium iodide (PI) had been analyzed by flow cytometry to figure out the percentage of cells displaying a rise in early (panels Q4) and late (panels Q2) apoptosis.PMID:23554582 The cells treated with camptothecin (15 mg/mL) and absolute ethanol (1 ; v/v) were made use of as constructive and damaging controls, respectively. Bar graphs show the summarized data on KKU-M213B (B), KKU-100 (D) and H69 (F) cells from two independent experiments per.