Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2008 Oct 21;36(20):6592–6607. doi: 10.1093/nar/gkn699

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© 2008 The Author(s)

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

PMC Copyright notice

Figure 5. — KDM1 inhibition by tranylcypromine and siRNA. (A) FACS analysis of HCT116 cells untreated and treated with tranylcypromine with and without nocodazole. (B) Q-PCR analysis of mRNA expression, in cells treated with tranylcypromine and/or nocodazole. The genes analyzed were Cyclin B1, Cyclin B2, CDC25C (G2/M) and PCNA (G1/S). All data were normalized for GAPDH expression and the non-cell-cycle regulated APOBEC3B was used as a further control. (C) ChIP analysis of H3/H4 methylations and of KDM1 and coREST binding along the Cyclin B2 and PCNA promoters. ChIPs were performed with the indicated antibodies on chromatin of cells arrested in G2/M, with and without tranylcypromine; black bars correspond to G2/M cells, white ones to G2/M cells treated with tranylcypromine. The values are normalized for the amount of H3/H4 immunoprecipitated in parallel. (D) Western blot of HCT116 nuclear extracts of untreated, tranylcypromine and G2/M-tranylcypromine-treated cells with the indicated antibodies. (E) Western blot analysis of HCT116 cells after KDM1 and coREST siRNA interference. Protein levels were compared with equal amounts of nuclear extracts derived from cells transfected with a control scramble siRNA. Nuclear extracts were prepared 40 h after transfections of siRNAs. (F) Gene expression analysis of cell-cycle genes after KDM1 and coREST RNA interference and nocodazole treatment, as in (E). Two different time points were checked in Q-PCR: 40 and 64 h after transfection. All data were GAPDH normalized. APOBEC3B gene was used as a further control.