bt549 cells  (MedChemExpress)

Journal: bioRxiv

Article Title: MYC Overexpression Confers Sensitivity to TACC3 Inhibition for Triple-Negative Breast Cancer

doi: 10.64898/2026.04.28.721171

Figure Lengend Snippet: a , Representative cell cycle profiles of RPE MYC and BT549 cells after treatment with BO-264 for 72 h. The percentage of cells in G1, S, and G2-M phases of the cell cycle, as determined by DNA content based on propidium iodide staining, is indicated. b , Flow cytometry gating strategy to identify GFP-positive cells (ISRE-active cells). Representative experimental control sample, untransduced BT549 cells. Side scatter and forward scatter were used to exclude debris. Forward scatter was used to distinguish single cells. Live cells were identified as negative for live/dead staining. GFP-positive live cells were identified by setting the fluorescence threshold based on untransduced BT549 controls. For reference, GFP signal intensity from live BT549 ISRE-GFP cells treated with IFNγ is shown to the left.

Article Snippet: BT549 cells were obtained from the American Type Culture Collection (ATCC) and were grown according to published guidelines .

Techniques: Staining, Flow Cytometry, Control, Fluorescence

Journal: bioRxiv

Article Title: MYC Overexpression Confers Sensitivity to TACC3 Inhibition for Triple-Negative Breast Cancer

doi: 10.64898/2026.04.28.721171

Figure Lengend Snippet: a , Immunoblot analysis of TACC3 expression in MTB/TOM and RPE cells. Representative image from n = 3 independent experiments. b , Immunoblot validation of siRNA knockdown of TACC3 in MYC OFF and MYC ON MTB/TOM cells (left) and RPE CONTROL and RPE MYC cells (right). Representative image from n = 3 independent experiments. c , Growth rate inhibition measurements of RPE CONTROL and RPE MYC cells in response to 72 h BO-264 treatment. Data show a representative experiment with n = 4 technical replicates. Error bars are mean ± s.d., and p values calculated using a two-sided t -test. Trends repeated across three independent cell passages. d , Crystal violet staining of RPE CONTROL and RPE MYC cells after 9 d treatment with increasing concentrations of BO-264. Images are representative of n = 3 independent experiments with similar results. e , Quantification of crystal violet stained RPE CONTROL and RPE MYC cells in response to various doses of BO-264. Error bars are mean ± s.e.m. P value was calculated using a two-sided t -test; * P < 0.05, *** P < 0.001, **** P < 0.0001. f , Immunoblot analysis of TACC3 proteasome-mediated degradation in RPE MYC (left) and BT549 (right) cells treated with BO-264 for 72 h. Prior to harvesting, cells were additionally incubated in the presence or absence of MG132. GAPDH is shown as a loading control. Representative image from n = 3 independent experiments.

Article Snippet: BT549 cells were obtained from the American Type Culture Collection (ATCC) and were grown according to published guidelines .

Techniques: Western Blot, Expressing, Biomarker Discovery, Knockdown, Control, Inhibition, Staining, Incubation

Journal: bioRxiv

Article Title: MYC Overexpression Confers Sensitivity to TACC3 Inhibition for Triple-Negative Breast Cancer

doi: 10.64898/2026.04.28.721171

Figure Lengend Snippet: a , Representative image of RPE MYC cells treated with 167nM BO-264 for 72 h with DAPI staining DNA. Arrows indicate micronuclei. Higher magnification of RPE MYC cells positive for micronuclei shown below. Scale bars 10uM unless indicated otherwise. b-c , Percentage of micronucleated RPE CONTROL , RPE MYC , BT549, and 4T1 cells 72 h after treatment with vehicle or BO-264 ( b ) or transfection with non-targeting (NT) or TACC3 siRNA ( c ). Data is an average of multiple high-powered (20x) fields analyzed per sample (700-1000 total cells/sample). Data representative of n = 3 independent experiments. Error bars are mean ± s.e.m., and p value calculated using a two-sided t-test.

Article Snippet: BT549 cells were obtained from the American Type Culture Collection (ATCC) and were grown according to published guidelines .

Techniques: Staining, Control, Transfection

Journal: bioRxiv

Article Title: MYC Overexpression Confers Sensitivity to TACC3 Inhibition for Triple-Negative Breast Cancer

doi: 10.64898/2026.04.28.721171

Figure Lengend Snippet: a , Flow cytometry cell cycle analysis with PI staining in RPE CONTROL and RPE MYC cells transfected with TACC3 siRNA (left) or treated with 167nM BO-264 (right) for 72 h. b , Immunoblot analysis of G1/S progression markers in RPE CONTROL and RPE MYC cells transfected with TACC3 siRNA (left) or treated with 167nM BO-264 (right) for 72 h. β-actin is shown as a loading control. Representative image from n = 3 independent experiments. c , Flow cytometry cell cycle analysis with PI staining in MYC OFF and MYC ON MTB/TOM cells treated with 300nM BO-264 for 72 h. d , Immunoblot analysis of mitotic arrest markers in MYC OFF and MYC ON MTB/TOM cells treated with 300nM BO-264 for 72 h. Representative image from n = 3 independent experiments. e , Flow cytometry cell cycle analysis with PI staining in BT549 cells treated with 300nM BO-264 for 72 h. f , Immunoblot analysis of mitotic arrest markers in BT549 cells treated with 300nM BO-264 for 72 h. Representative image from n = 3 independent experiments. Data representative of n = 3 independent experiments. Error bars are mean ± s.e.m., and p values calculated using a two-sided t -test.

Article Snippet: BT549 cells were obtained from the American Type Culture Collection (ATCC) and were grown according to published guidelines .

Techniques: Flow Cytometry, Cell Cycle Assay, Staining, Control, Transfection, Western Blot

Journal: bioRxiv

Article Title: MYC Overexpression Confers Sensitivity to TACC3 Inhibition for Triple-Negative Breast Cancer

doi: 10.64898/2026.04.28.721171

Figure Lengend Snippet: a , Diagram of IFN induction assay: BT549 cells contain an IFN reporter expressing destabilized GFP (dscGFP). ISRE: IFN-stimulated response elements. GFP median fluorescence intensity (MFI) was assessed 72 h after treatment by flow cytometry. Created with BioRender.com. b , Representative flow cytometry plot depicting GFP MFI of BT549 ISRE-GFP cells treated with vehicle, BO-264, or IFNγ for 72 h. Untransduced BT549 cells were used to establish GFP-positivity. c , Quantification of GFP positivity in BT549 ISRE-GFP cells after 72 h treatment. d , Flow cytometry analysis of MHC MFI in RPE (left) and 4T1 (right) cells treated with BO-264 for 72 h. Representative experiment with three samples per condition shown. Error bars are mean ± s.d., and p values calculated using a two-sided t -test. Trends repeated across three independent cell passages. ( e - f ), ELISA analysis of secreted CCL2 ( e ) and CXCL10 ( f ) levels in the indicated cell lines following treatment with vehicle or BO-264 for 72 h. g , ELISA analysis of intracellular cGAMP levels in the indicated cell lines upon treatment with vehicle or BO-264 for 72 h. h , Immunoblot analysis of cGAS-STING pathway activity in MTB/TOM MYC ON , 4T1, BT549, and RPE MYC cells treated with BO-264 for 72 h. β-actin is shown as a loading control. Representative image from n = 3 independent experiments. Data representative of n = 3 independent experiments. Error bars are mean ± s.e.m., and P values calculated using a two-sided t -test.

Article Snippet: BT549 cells were obtained from the American Type Culture Collection (ATCC) and were grown according to published guidelines .

Techniques: Expressing, Fluorescence, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Western Blot, Activity Assay, Control

Journal: bioRxiv

Article Title: MYC Overexpression Confers Sensitivity to TACC3 Inhibition for Triple-Negative Breast Cancer

doi: 10.64898/2026.04.28.721171

Figure Lengend Snippet: a , Flow cytometry analysis of DC activation (HLA-DR + CD86 + ) across six donors following DC co-culture with conditioned medium from BT549 cells treated with indicated drugs for 72h. Data are representative of n = 3 technical replicates per donor. Error bars indicate mean ± s.d., and P values were calculated using a two-sided t -test.

Article Snippet: BT549 cells were obtained from the American Type Culture Collection (ATCC) and were grown according to published guidelines .

Techniques: Flow Cytometry, Activation Assay, Co-Culture Assay