Journal: Cell Death & Disease

Article Title: Dual targeting of PI3Kδ and PPARα enhances antitumor activity via FoxO1 activation in follicular lymphoma

doi: 10.1038/s41419-026-08593-5

Figure Lengend Snippet: A - C EdU flow plots for Karpas-422, RL, and Sc-1 cells after 24-h treatment (Linperlisib 32 μM; Chiglitazar 16 μM). D – F Quantification of EdU-positive cells showing reduced DNA synthesis with the combination ( n = 3). G – L ( G , H ) for Karpas-422; ( I , J ) for RL; ( K , L ) for Sc-1. Cell-cycle distribution by PI staining, demonstrating G0/G1 accumulation with Linperlisib and further enhancement by the combination (Linperlisib 16 μM; Chiglitazar 8 μM; n = 3). M GSEA indicating negative enrichment of the G1/S transition pathway (NES = − 2.85865, p < 0.00001). N – P Western blots of G1/S checkpoint proteins (p27, Cyclin E1, CDK2, p-CDK2); combination treatment increased p27 and decreased Cyclin E1, CDK2, and p-CDK2 (representative of three independent experiments).

Article Snippet: The primary antibodies used were as follows: PI3K p110δ (A19742; ABclonal), pan-Akt (#4691; CST), phospho-Akt (Ser473, #4060; CST), PPARα (ab227074; Abcam), FoxO1 (#2880; CST), phospho-FoxO1 (Thr24, #9464; CST) and (Ser256, #9461; CST), FoxO3a (#12829; CST), FoxO4 (#9472; CST), Mcl-1 (#16225-1-AP; Proteintech), Bcl-2 (#12789-1-AP; Proteintech), Bim (A19702; ABclonal), Bax (#50599-2-Ig; Proteintech), Cleaved PARP(#5625; CST), P21(#2947; CST), p27 (#3686; CST), Cyclin E1 (#11935-1-AP; Proteintech), CDK2 (#10122-1-AP; Proteintech), phospho-CDK2 (#4539; CST), GLUT1(#73015; CST), PGK1(A12686; ABclonal), β-Tubulin (#10094-1-AP; Proteintech), Lamin B1 (#13435; CST), and β-Actin (#66009-1-Ig; Proteintech).

Techniques: DNA Synthesis, Staining, Western Blot

Journal: FEBS Open Bio

Article Title: Chemical and molecular bases of dome formation in human colorectal cancer cells mediated by sulphur compounds from Cucumis melo var. conomon

doi: 10.1002/2211-5463.13001

Figure Lengend Snippet: Enriched GO terms related to cell‐cycle control for the genes down‐regulated by MTA.

Article Snippet: The membrane was then sequentially incubated with the following solutions at room temperature: 5% skim milk in PBS containing 0.1% Tween 20 for 1 h; the primary antibodies against human CCNE2 (1 : 2000, RRID: AB_1847388; Merck KGaA), human CDC25A (1 : 100, RRID: AB_627226; Santa Cruz Biotechnology, Inc., Dallas, TX, USA) or chicken actin (1 : 2000, RRID: AB_63314; Life Technologies) for 2 h; and the horseradish peroxidase‐conjugated secondary antibodies (1 : 1500–2500) against anti‐chicken IgY for CCNE2 detection (RRID: AB_228385; Life Technologies), and anti‐mouse IgG for CDC25A and actin detection (RRID: AB_2687537; SeraCare Life Sciences, Inc., Milford, MA, USA) for 1 h. The immunoreactive CCNE2 protein was visualized on X‐ray film using the enhanced chemiluminescence detection system (GE Healthcare).

Techniques: Control

Journal: FEBS Open Bio

Article Title: Chemical and molecular bases of dome formation in human colorectal cancer cells mediated by sulphur compounds from Cucumis melo var. conomon

doi: 10.1002/2211-5463.13001

Figure Lengend Snippet: MTA down‐regulated genes related to cell‐cycle control.

Article Snippet: The membrane was then sequentially incubated with the following solutions at room temperature: 5% skim milk in PBS containing 0.1% Tween 20 for 1 h; the primary antibodies against human CCNE2 (1 : 2000, RRID: AB_1847388; Merck KGaA), human CDC25A (1 : 100, RRID: AB_627226; Santa Cruz Biotechnology, Inc., Dallas, TX, USA) or chicken actin (1 : 2000, RRID: AB_63314; Life Technologies) for 2 h; and the horseradish peroxidase‐conjugated secondary antibodies (1 : 1500–2500) against anti‐chicken IgY for CCNE2 detection (RRID: AB_228385; Life Technologies), and anti‐mouse IgG for CDC25A and actin detection (RRID: AB_2687537; SeraCare Life Sciences, Inc., Milford, MA, USA) for 1 h. The immunoreactive CCNE2 protein was visualized on X‐ray film using the enhanced chemiluminescence detection system (GE Healthcare).

Techniques: Control

Journal: FEBS Open Bio

Article Title: Chemical and molecular bases of dome formation in human colorectal cancer cells mediated by sulphur compounds from Cucumis melo var. conomon

doi: 10.1002/2211-5463.13001

Figure Lengend Snippet: RT‐PCR verification of the MTA‐mediated down‐regulation of CCNE2 and CDC25A . RT‐PCR was performed to confirm the reduction in CCNE2 and CDC25A mRNA expression that was screened in the microarray analysis. After MTA treatment for the indicated hours, the total RNA was subjected to RT‐PCR. The electrophoretically separated PCR bands were stained with ethidium bromide, and representative gel images (A) of three replicates are shown. The numbers under the photographs show the fluorescence intensity of each band relative to an initial value (= 1.00), which was calculated after normalization against GAPDH as a reference. The bar graphs show the fluctuations of the CCNE2 (B) and CDC25A (C) expression levels obtained independently by the RT‐PCR and microarray analyses. The relative mRNA levels determined by RT‐PCR represent mean ± SD of three experiments. Values with different letters are significantly different by one‐way ANOVA followed by Tukey–Kramer HSD test ( P < 0.01). The characters in parentheses indicate the probe set ID of the microarray.

Article Snippet: The membrane was then sequentially incubated with the following solutions at room temperature: 5% skim milk in PBS containing 0.1% Tween 20 for 1 h; the primary antibodies against human CCNE2 (1 : 2000, RRID: AB_1847388; Merck KGaA), human CDC25A (1 : 100, RRID: AB_627226; Santa Cruz Biotechnology, Inc., Dallas, TX, USA) or chicken actin (1 : 2000, RRID: AB_63314; Life Technologies) for 2 h; and the horseradish peroxidase‐conjugated secondary antibodies (1 : 1500–2500) against anti‐chicken IgY for CCNE2 detection (RRID: AB_228385; Life Technologies), and anti‐mouse IgG for CDC25A and actin detection (RRID: AB_2687537; SeraCare Life Sciences, Inc., Milford, MA, USA) for 1 h. The immunoreactive CCNE2 protein was visualized on X‐ray film using the enhanced chemiluminescence detection system (GE Healthcare).

Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, Microarray, Staining, Fluorescence

Journal: FEBS Open Bio

Article Title: Chemical and molecular bases of dome formation in human colorectal cancer cells mediated by sulphur compounds from Cucumis melo var. conomon

doi: 10.1002/2211-5463.13001

Figure Lengend Snippet: MTA‐induced reduction of the CCNE2 and CDC25A proteins in RCM‐1 cells. Following MTA treatment for the indicated hours, proteins extracted from the cells were applied to immunoblotting with the respective amounts of 3.75 µg (per each lane) for detecting CCNE2 (A) and 30 µg for CDC25A (B). The photographs show the immunologically detected proteins together with actin as a reference.

Article Snippet: The membrane was then sequentially incubated with the following solutions at room temperature: 5% skim milk in PBS containing 0.1% Tween 20 for 1 h; the primary antibodies against human CCNE2 (1 : 2000, RRID: AB_1847388; Merck KGaA), human CDC25A (1 : 100, RRID: AB_627226; Santa Cruz Biotechnology, Inc., Dallas, TX, USA) or chicken actin (1 : 2000, RRID: AB_63314; Life Technologies) for 2 h; and the horseradish peroxidase‐conjugated secondary antibodies (1 : 1500–2500) against anti‐chicken IgY for CCNE2 detection (RRID: AB_228385; Life Technologies), and anti‐mouse IgG for CDC25A and actin detection (RRID: AB_2687537; SeraCare Life Sciences, Inc., Milford, MA, USA) for 1 h. The immunoreactive CCNE2 protein was visualized on X‐ray film using the enhanced chemiluminescence detection system (GE Healthcare).

Techniques: Western Blot

Journal: FEBS Open Bio

Article Title: Chemical and molecular bases of dome formation in human colorectal cancer cells mediated by sulphur compounds from Cucumis melo var. conomon

doi: 10.1002/2211-5463.13001

Figure Lengend Snippet: CCNE2 accumulation in various mammalian cells. Immunoblotting was performed to examine CCNE2 accumulation in different nontransformed and transformed mammalian cell lines. The following cell lines were used: WB‐F344, WB‐F344 rat liver epithelial cells; WB‐Ha‐ras, v‐Ha‐ras‐transformed WB; WB‐neu, neu‐transformed WB; WBsrc‐neo2, v‐src‐transformed WB; WB‐myc/ras, myc/ras‐transformed WB; Ehrlich, Ehrlich ascite tumor cells; HPD1NR, transplantable pancreatic ductal cancer cells; HaPT1, hamster pancreatic cancer cells; MSU‐2, human foreskin fibroblast cells; KNC, human well‐differentiated colon cancer cells; RCM‐1, human well‐differentiated rectal cancer cells; CoCM‐1, human moderately differentiated colon cancer cells; HCT116, human poorly differentiated colon cancer cells; BxPC3 and Ming, human pancreatic cancer cells.

Article Snippet: The membrane was then sequentially incubated with the following solutions at room temperature: 5% skim milk in PBS containing 0.1% Tween 20 for 1 h; the primary antibodies against human CCNE2 (1 : 2000, RRID: AB_1847388; Merck KGaA), human CDC25A (1 : 100, RRID: AB_627226; Santa Cruz Biotechnology, Inc., Dallas, TX, USA) or chicken actin (1 : 2000, RRID: AB_63314; Life Technologies) for 2 h; and the horseradish peroxidase‐conjugated secondary antibodies (1 : 1500–2500) against anti‐chicken IgY for CCNE2 detection (RRID: AB_228385; Life Technologies), and anti‐mouse IgG for CDC25A and actin detection (RRID: AB_2687537; SeraCare Life Sciences, Inc., Milford, MA, USA) for 1 h. The immunoreactive CCNE2 protein was visualized on X‐ray film using the enhanced chemiluminescence detection system (GE Healthcare).

Techniques: Western Blot, Transformation Assay