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. 2012 May 21;109(23):8971–8976. doi: 10.1073/pnas.1119836109

Fig. 3.

Fig. 3.

Jarid1a associates with RB and induces premature senescence. (A) Immunoblots from chromatin-bound fractions of growing (G), quiescent (Q), and senescent (S) cells using antibodies that specifically recognize Jarid1a or Jarid1b. Coomassie blue staining of core histone proteins is shown as evidence of equal loading. (B) Immunoblots showing Jarid1a coimmunoprecipitation with RB. Note the specific Jarid1a signal in the in the senescent immunoprecipitation, but those from growing and quiescent immunoprecipitates show a nonspecific lower molecular weight smear. (C) Immunoblots documenting expression of Jarid1a, levels of H3K4me3, and levels of MCM3 and Cyclin A in ras-senescent (S), empty vector (V) infected, and Jarid1a-infected IMR90 cells. Parallel blots were probed with an anti-Actin antibody as a protein quantification control. (D) Micrographs showing the senescence markers SA-β-gal and SAHF in Jarid1a-infected IMR90 cells. (E) Immunoblots documenting expression of wild-type or catalytic inactive mutant (H483A) Jarid1a, levels of H3K4me3, and levels of MCM3, Cyclin A, and PCNA. Parallel blots were probed with an anti-Actin antibody as a protein quantification control. (F) Quantification of senescence markers. Values represent the ± SE of at least three independent experiments.