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Fig.1 | Epigenetics & Chromatin

Fig.1

From: The MMP-2 histone H3 N-terminal tail protease is selectively targeted to the transcription start sites of active genes

Fig.1

The MMP-2 protease directs H3NT proteolysis in U2OS cells. a Illustration of experimental time points examined in this study based on U2OS cell density, ranging from subconfluent (− 2 days) to confluent (0) to overconfluent (+ 2 days). b A histone H3 C-terminal antibody was used for Western analysis of chromatin purified from U2OS cells at the indicated time points. Proteolysis of the H3 N-terminus (H3NT) generates faster-migrating cleaved H3 (H3cl) products as indicated. Amido black stain (AB) of the membrane shows equivalent loading of chromatin between samples. Lower panel shows Western analysis of purified soluble nuclear extracts at these time points using an MMP-2 antibody. c Western analysis of purified chromatin from 293 T negative control cells, C2C12 differentiated myotube positive control, and U2OS + 2 and − 2 cells. The MMP-2 generated H3NT cleaved product (H3∆18) and two other H3 cleaved products, a slower migrating product (H3cl.s) and a faster migrating product (H3cl.f), are indicated. d Western analysis of nuclear extracts purified from stable U2OS + 2 cell lines expressing the pLKO.1 negative control or pLKO.1-MMP-2 shRNAs (sh1 and sh2) demonstrates depletion of the pro-form and catalytically active form of MMP-2. e Western analysis of chromatin from the same samples as in d

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