Journal: Neoplasia (New York, N.Y.)

Article Title: GBP1-CDK9-STAT3 signaling axis promotes osteosarcoma PD-L1 expression and immune escape

doi: 10.1016/j.neo.2025.101232

Figure Lengend Snippet: GBP1 drives STAT3 activation and PD-L1 upregulation through CDK9.

Article Snippet: The STAT3 inhibitor STAT3-IN-13 (HY-150603) and the CDK9 inhibitor LDC000067 (Synonyms: LDC067, HY-15878) were purchased from MedChemExpress (Monmouth Junction, USA). siRNAs were procured from RiboBio (Guangzhou, China).

Techniques: Activation Assay

Journal: Cancer Medicine

Article Title: miR‐206 inhibits the growth of hepatocellular carcinoma cells via targeting CDK9

doi: 10.1002/cam4.1188

Figure Lengend Snippet: Overexpression of miR‐206, shCDK9, and LDC000067 suppresses the expression of CDK9 and its downstream genes. Bell7402 and HepG2 cells were transfected with GV251‐miRNA‐206 (A, E), sh‐CDK9 (C, D and F), and screened with G418 for 2–4 weeks to establish the stable cell lines or treated with 15 μmol/L LDC000067 (G) for 48 h. Then the levels of CDK9 mRNA were assayed by qPCR (A, C) taking β‐actin as a control, and the protein levels of CDK9, p‐RNAII, t‐RNAII, and MCL‐1 were detected by western blot (B, D–G) taking tubulin as the loading control. * P < 0.05.

Article Snippet: LDC000067 (CDK9 inhibitor) was purchased from Amquar Company (Shanghai, China). sh‐CDK9 plasmid was synthesized by GeneChem Company (Shanghai, China).

Techniques: Over Expression, Expressing, Transfection, Stable Transfection, Control, Western Blot

Journal: Cancer Medicine

Article Title: miR‐206 inhibits the growth of hepatocellular carcinoma cells via targeting CDK9

doi: 10.1002/cam4.1188

Figure Lengend Snippet: Inhibition of CDK9 by shCDK9 and LDC000067 represses the cell proliferation and induces apoptosis and cell cycle arrest in HCC cell lines. Bell7402 and HepG2 cells were transfected with sh‐CDK9 and screened with G418 for 2–4 weeks to establish the stable cell lines or treated with 15 μmol/L LDC000067 for 48 h, then the cell proliferation (A and B) was examined by CCK‐8 assays. The apoptosis (C–J) and the cell cycle phase distribution (K–L) were detected by flow cytometry. * P < 0.05.

Article Snippet: LDC000067 (CDK9 inhibitor) was purchased from Amquar Company (Shanghai, China). sh‐CDK9 plasmid was synthesized by GeneChem Company (Shanghai, China).

Techniques: Inhibition, Transfection, Stable Transfection, CCK-8 Assay, Flow Cytometry

Journal: Developmental cell

Article Title: Quiescent cells actively replenish CENP-A nucleosomes to maintain centromere identity and proliferative potential

doi: 10.1016/j.devcel.2019.07.016

Figure Lengend Snippet: (A) Single molecule RNA fish showing the presence of centromeric alpha satellite-derived transcripts in RPE-1 cells. Scale bar = 5 μm. (B) Quantification of transcript numbers. Transcripts are approximately equal in cycling and G0 quiescent cells, but decrease when treated with the Cdk7 transcription inhibitor THZ1. Error bars represent the mean and standard deviation (cycling n = 267 nuclei; cycling + THZ1 n = 113; quiescent n = 269; quiescent + THZ1 n = 183). ****p<0.0001 by Mann Whitney test. (C) Immunofluorescence for RNA Pol II and detection of nascent RNA synthesis by EU incorporation reveals ongoing transcription in prophase I arrested starfish oocytes. Scale bar = 5 μm. (D) Pol II and nascent RNA (EU) are present in the vicinity of centromeres, visualized with 3xGFP-CENP-N in arrested oocytes. Pol II and EU images are scaled equivalently. (E) Treatment with the Pol II inhibitor Triptolide reduces Pol II localization and EU levels at centromeres. Individual points represent the average fluorescence at all centromeres. Error bars represent the mean and standard deviation (control n = 51 oocytes, triptolide n = 57 oocytes, measured for both Pol II and EU). ****p<0.0001 by Mann Whitney test. (F) Immunofluorescence showing new CENP-A incorporation in quiescent oocytes. Inhibition of Pol II with 10 μM of triptolide, or Cdk9 with 1 μM of LDC000067 reduces incorporation of new 3xGFP-CENP-A in prophase after 9 days in culture. Incorporation occurs in prophase I, but oocytes were stimulated to enter meiosis I for visualization to condense chromosomes for clarity. Scale bar = 5 μm. Metaphase 3xGFP-CENP-A images are scaled equivalently, and prophase images are linearly scaled individually (G) Quantification of new 3xGFP-CENP-A incorporation relative to DMSO control. Each point represents the mean of all centromeres from one oocyte. Error bars represent the mean and standard deviation (control, n = 58; triptolide, n = 47; Cdk9i, n = 51). ****p<0.0001 by Mann Whitney test. (H) Inhibition with 10 μM triptolide or 1 μM LDC000067 during prophase I arrest and in meiosis does not alter incorporation of new 3xGFP-CENP-A in G1. Scale bars = 5 μm. (I) Quantification of 3xGFP-CENP-A incorporation relative to DMSO control. Each point represents the mean of all centromeres from one oocyte. Error bars represent the mean and standard deviation (control n = 42, triptolide n = 20, Cdk9i n = 38). Significance determined by Mann Whitney test.

Article Snippet: For transcription studies , 10 μM triptolide (Pol II inhibitor, Sigma) or 1 μM LDC000067 (Cdk9 inhibitor, Selleck Chemicals) was added to CF and changed every 2 days.

Techniques: Derivative Assay, Standard Deviation, MANN-WHITNEY, Immunofluorescence, Fluorescence, Inhibition