Journal: Translational Oncology

Article Title: The novel tertiary amine LSD1 inhibitor 596 inhibits endometrial cancer through the mTOR signal transduction pathway

doi: 10.1016/j.tranon.2026.102688

Figure Lengend Snippet: 596 interacts with LSD1 in EC Cells. (A). The molecular structure of 596. (B). Western blot detection of H3K4me1, H3K4me2, and H3K4me3 after 596(0,4,8,12,16 μM) treatment( n = 3). (C-D). After treating ISK and HEC1A cells with DMSO and 596 (12 μM) for 2 h, the thermal stability of LSD1 was detected by CETSA method. The quantitation (right) of western blot results. (E). In ISK and HEC1A cells, detect the proliferation of compound 596 after 48–96 h of different concentrations by CCK8. (F-G). GraphPad Prism 9 was used to calculate IC50 of 596 in ISK and HEC1A cells. (H). ISK cells and HEC1A cells were treated with different concentrations of 596 respectively, and the proliferation of the cells was detected by colony formation assay.

Article Snippet: The EC cells were cultured in DMEM medium(Viva cell, China) containing 10% fetal bovine serum (Viva cell, China) at 37°C and 5% CO 2 .The main purchasing information was listed below:LSD1 (1:10,000, ab129195, Abcam, UK), H3(1:20,000, No.4499T,Cell Signaling Technology, CST, Boston, MA, USA), H3K4me1(1:2000, No.5326T, CST), H3K4me2(1:2000, No.9725S, CST), H3K4me3(1:2000, No. 9751T, CST), p62/SQSTM1(1:2000, No.5114T, CST), LC3B(1:2000, Cat# AF4650, Affinity Biosciences), Beclin-1(1:2000, Cat# AF5128, Affinity Biosciences), AKT(1:5000, YM8463, Immunoway, USK), P-AKT(Ser473)(1:2000, YM8304, Immunoway, USK), mTOR(1:5000,YM8208, Immunoway, USK), P-mTOR(Ser2448)(1:5000, YM8326, Immunoway, USK), S6(1:2000, YT4139, Immunoway, USK), P-S6(1:2000, No.F0198, Selleck.cn), p70S6K(1:2000, YT3559, Immunoway, USK), P-p70S6K(1:1000, No.F2907, Immunoway, USK), GAPDH(1:5000, Proteintech).

Techniques: Western Blot, Quantitation Assay, Colony Assay