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. 2020 Mar 10;9:e51373. doi: 10.7554/eLife.51373

Figure 1. PHF19L associates with PRC2 in prostate cancer cells.

(A) Schematic representation of PHF19L and PHF19S and their domains. (B) Western blot analysis showing expression of PHF19L, PHF19S, EZH2, and GAPDH in RWPE1, PC3, and DU145 cells. *, non-specific bands (C) Summary of the main interactors of PHF19L and PHF19S identified by mass spectrometry (MS). PC3 cells stably expressing FLAG-tagged PHF19L or PHF19S, or FLAG-tagged empty vector (control), were subjected to FLAG affinity purification followed by MS. The table displays the score and the peptide count from two independent experiments. (D,E) Endogenous co-immunoprecipitation (IP) of PHF19L with EZH2 or SUZ12 in control (shCTR) and PHF19L-depleted (shPHF19L#1 or shPHF19L#4) PC3 cells (D) or DU145 cells (E). IgG was used as a control. *, non-specific band.

Figure 1.

Figure 1—figure supplement 1. PHF19L associates with PRC2 in prostate cancer cells.

Figure 1—figure supplement 1.

(A) Co-immunoprecipitation (IP) of FLAG-tagged proteins in PC3 cells overexpressing FLAG-PHF19L, FLAG-PHF19S, or FLAG-Empty. IP experiments were performed using anti FLAG-M2 antibody followed by EZH2 and PHF19L detection by Western blot. (B) RT-qPCR showing expression of the PHF19L (upper panels) and PHF19S (lower panels) isoforms in PC3 (left panels) and DU145 (right panels) cells. Cells were transduced with lentivirus expressing different shRNAs against PHF19L (shPHF19L#1, shPHF19L#4 or shPHF19L#B), PHF19S (shPHF19S#55 or shPHF19S#168) or a control shRNA (shCTR). Results are shown relative to shCTR and are normalized to the housekeeping gene RPLPO. Data represent mean ± SD from three biological replicates.