mcf 10a cell lines  (ATCC)

Journal: Oncology Letters

Article Title: Protein lactylation within the nucleus independently predicts the prognosis of non‑specific triple‑negative breast cancer

doi: 10.3892/ol.2024.14818

Figure Lengend Snippet: Oxamate decreased Klac levels and inhibited the proliferation of MDA-MB-231 cells. (A) Elevated global Klac levels in the triple-negative breast cancer cell line, MDA-MB-231 compared with the benign mammary epithelial cell line, MCF-10A (loading control: β-actin; whole cell lysate: 20 µg). (B) Significantly repressed cell proliferation after oxamate (20 mM) treatment in MDA-MB-231 cells (blue: control group; red: oxamate group). (C) Significantly decreased global Klac levels after oxamate treatment (loading control: β-actin; whole cell lysate: 20 µg). (D) Significantly decreased Klac levels of histones after oxamate treatment (loading control: β-actin; for Klac: whole cell lysate: 30 µg). The graph is the semi-quantified data of the western blot bands (n=3). (E) Dose-dependent reductions in lactylation levels at H3, H3K14 and H3K18 residues after oxamate treatment (loading control: H3). The analysis was conducted using histone protein extract (2 µg). The graph is the semi-quantified data of the western blot bands (n=3). The membranes were cut in accordance with the molecular weight markers electrophoresed on one or both sides of the membrane before its hybridization with antibodies. All experiments were repeated three times. Statistical significance between the indicated groups in the cell viability and western blotting assays was assessed with the two-tailed unpaired Student's t-test. *P<0.05, **P<0.01. Ct, control group; Oxa, oxamate group; Klac, lysine lactylation.

Article Snippet: The MCF-10A cells were maintained in Dulbecco's Modified Eagle Medium/F12 (1:1) supplemented with 10% FBS, 10 µg/ml insulin, 20 ng/ml epidermal growth factor, 100 U/ml penicillin and 100 µg/ml streptomycin (cat. no. CL-0525; Procell Life Science & Technology Co., Ltd.).

Techniques: Control, Western Blot, Molecular Weight, Membrane, Hybridization, Two Tailed Test

Journal: Oncology Letters

Article Title: Protein lactylation within the nucleus independently predicts the prognosis of non‑specific triple‑negative breast cancer

doi: 10.3892/ol.2024.14818

Figure Lengend Snippet: Oxamate decreased Klac levels and inhibited the proliferation of MDA-MB-231 cells. (A) Elevated global Klac levels in the triple-negative breast cancer cell line, MDA-MB-231 compared with the benign mammary epithelial cell line, MCF-10A (loading control: β-actin; whole cell lysate: 20 µg). (B) Significantly repressed cell proliferation after oxamate (20 mM) treatment in MDA-MB-231 cells (blue: control group; red: oxamate group). (C) Significantly decreased global Klac levels after oxamate treatment (loading control: β-actin; whole cell lysate: 20 µg). (D) Significantly decreased Klac levels of histones after oxamate treatment (loading control: β-actin; for Klac: whole cell lysate: 30 µg). The graph is the semi-quantified data of the western blot bands (n=3). (E) Dose-dependent reductions in lactylation levels at H3, H3K14 and H3K18 residues after oxamate treatment (loading control: H3). The analysis was conducted using histone protein extract (2 µg). The graph is the semi-quantified data of the western blot bands (n=3). The membranes were cut in accordance with the molecular weight markers electrophoresed on one or both sides of the membrane before its hybridization with antibodies. All experiments were repeated three times. Statistical significance between the indicated groups in the cell viability and western blotting assays was assessed with the two-tailed unpaired Student's t-test. *P<0.05, **P<0.01. Ct, control group; Oxa, oxamate group; Klac, lysine lactylation.

Article Snippet: The human TNBC cell line, MDA-MB-231, and the human normal mammary ductal cell line, MCF-10A, were provided by Procell Life Science & Technology Co., Ltd.

Techniques: Control, Western Blot, Molecular Weight, Membrane, Hybridization, Two Tailed Test

Journal: Nature Communications

Article Title: DCAF13-mediated K63-linked ubiquitination of RNA polymerase I promotes uncontrolled proliferation in Breast Cancer

doi: 10.1038/s41467-025-55851-9

Figure Lengend Snippet: a The top 10 BC prognosis-associated RiBPs from the TCGA-BC and Chia-Ho cohorts. The upper bar chart displaying the -log 10 p -value for these RiBPs, derived from Kaplan-Meier overall survival analysis. The middle and lower heatmap showing the p -value and associated log 2 FC for these RiBPs, respectively. b Violin plots illuminating the differential mRNA expression levels of DCAF13, DKC1, and RPP40 between BC and normal tissue samples. c Kaplan-Meier overall survival curves elucidating the prognostic significance of DCAF13, DKC1, and RPP40 mRNA expression levels in BC patients. d A SUnSET assay executed on MDA-MB-231 cells following the knockdown of each of the three aforementioned AMFs. On the right is the corresponding statistical graph ( n = 3 biological replicates). e, f CCK8 assay ( e ) and colony formation assay ( n = 3 biological replicates) ( f ) administered to MDA-MB-231 cells after knockdown of DCAF13, DKC1, and RPP40, respectively. g Colony formation assay performed on MCF10A cells following the knockdown of DCAF13, DKC1, and RPP40, respectively. The corresponding statistical graph is shown on the right ( n = 3 biological replicates). b Box plots show the distribution of data across groups. The central line within the box represents the median value. The upper and lower edges of the box represent the 75th and 25th percentiles, respectively (Interquartile Range, IQR). The whiskers extend from the box to the maximum and minimum values within 1.5 times the interquartile range (IQR), with any data points beyond this range considered as outliers. a – d, f, g Data shown as mean ± SEM. P -values of survival analysis were calculated using the two-sided Log-rank (Mantel-Cox) test, p -value of differential expression analysis were calculated using the Likelihood Ratio Test, and subsequently adjusted using the Benjamini-Hochberg (BH) method to obtain False Discovery Rate (FDR) value ( a ). Statistical significance was calculated by two-sided Wilcoxon rank-sum test ( b ), two-sided Log-rank (Mantel-Cox) test ( c ), two-tailed unpaired Student’s t-test ( d, f, g ). Source data are provided as a Source Data file.

Article Snippet: Human cell lines MDA-MB-468, MDA-MB-231, MCF10A, HMLE, HeLa, and HEK293Twere all procured from ATCC.

Techniques: Derivative Assay, Expressing, Knockdown, CCK-8 Assay, Colony Assay, Two Tailed Test