Journal: Oncology Reports
Article Title: KDM5D expression is lost in cisplatin-resistant neuroblastoma cells
doi: 10.3892/or.2026.9084
Figure Lengend Snippet: KDM5D and CDDP treatment affect expression of CUL4A in UKF-NB-3 and UKF-NB-3 CDDP. Flow cytometry analysis in (A) UKF-NB-3 and UKF-NB-3 CDDP , where sensitive cell line UKF-NB-3 showed lower expression of CUL4A gene compared with the resistant cell line (P<0.05) and this expression increased in sensitive cells after CDDP treatment (P<0.001) compared with control. (B) Silencing of KDM5D increased of CUL4A level (P<0.001). (C) In the CDDP-resistant cell line UKF-NB-3 CDDP , KDM5D overexpression decreased CUL4A expression compared with mock and control plasmid (P<0.001). CDDP treatment of overexpressed cells with KDM5D increased the level of CUL4A (P<0.001), while in control cells CDDP treatment did not change the level of CUL4A. Data are shown as mean ± standard deviation from three independent experiments. Statistical significance was determined using two-way ANOVA with Tukey's post hoc test. *P<0.05; **P<0.01; ***P<0.001 (ANOVA with Tukey's post hoc test). CTR, control; NC, non-coding RNA; si, short interfering RNA; pl, ORF-clone plasmid; NB3, UKF-NB-3; Mock/NC, control cells; siKDM5D/pl-KDM5D, control cells transfected with siKDM5D/pl-KDM5D; FC, flow cytometry.
Article Snippet: After washing, the cell pellets were blocked with 5% BSA in PBS for 1 hour at room temperature After blocking, the pellets were incubated with the primary antibody anti-JARID1D rabbit mAB at a dilution of 1:100 (cat. no. PA5-100844; Invitrogen, Thermo Fisher Scientific, Inc.), anti-JARID1B rabbit mAB at dilution of 1:400 (cat. no. 15327S; Cell Signaling Technology, Inc.), anti-trimethyl histone H3 (Lys4) rabbit (cat. no. 07-473; MilliporeSigma) at a dilution of 1:400, Cleaved-Caspase 3 (Asp175) Rabbit mAB at a dilution of 1:50 (cat. no. 9602S; Cell Signaling Technology, Inc.) or CUL4A Rabbit mAB at a dilution of 1:50 (cat. no 2699; Cell Signaling Technology, Inc.) for 1 h at laboratory temperature.
Techniques: Expressing, Flow Cytometry, Control, Over Expression, Plasmid Preparation, Standard Deviation, Small Interfering RNA, Transfection