cells  (MedChemExpress)

Journal: Advanced Science

Article Title: Nanoparticle‐Mediated TIPE1 mRNA Delivery Enhances Paclitaxel Sensitivity in Triple‐Negative Breast Cancer by Modulating RAB7A Ubiquitination‐Associated Stability and Autophagy

doi: 10.1002/advs.76975

Figure Lengend Snippet: TIPE1 modulates PTX‐induced cytotoxicity by regulating autophagy and mitochondrial function. (A) Experimental workflow illustrating the strategy for assessing changes in autophagy and mitochondrial function following TIPE1 knockdown in MDA‐MB‐231 cells and overexpression in MDA‐MB‐231R cells; (B,C) RT‐qPCR and WB analysis of Beclin‐1 and ATG5 mRNA expression following TIPE1 knockdown in MDA‐MB‐231 cells; (D,E) RT‐qPCR and WB analysis of Beclin‐1, ATG5, and p62 expression after oe‐TIPE1; (F) JC‐1 staining to evaluate changes in MMP following TIPE1 modulation (bar = 25 µm); (G) DCFH‐DA fluorescent probe to detect intracellular ROS levels, including Mito‐TEMPO pretreatment conditions (bar = 25 µm); (H) TUNEL staining to assess apoptosis levels after PTX treatment (bar = 50 µm). All data are presented as mean ± SD. Experiments were performed in triplicate. Statistical significance was determined by ANOVA followed by Tukey's post hoc test. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: For Mito‐TEMPO treatment, cells were pretreated with 5 μ m Mito‐TEMPO (HY‐112879, MedChemExpress, USA) for 1 h before PTX treatment for 48 h. Cells were then incubated with 10 μ m DCFH‐DA (D6883, Sigma–Aldrich, USA) at 37°C in the dark for 30 min and washed three times with PBS to remove extracellular probe.

Techniques: Knockdown, Over Expression, Quantitative RT-PCR, Expressing, Staining, TUNEL Assay