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. 2017 Jun 12;9(8):1030–1051. doi: 10.15252/emmm.201607088

Figure EV5. Analysis of subcellular localization and function of E2F5 in HPV(+) HNSCC cells.

Figure EV5

  1. Effects of shRNA‐mediated knockdown on E2F5 on ATG5, Drp1, and LC3B mRNAs were measured using qRT–PCR in UM‐SCC‐47 cells compared to Scr‐shRNA‐transfected controls. Data are means ± SD from three independent experiments.
  2. Subcellular localization of E2F5 was assessed in the presence/absence of C18‐pyr‐cer (20 μM, 1 h) by immunofluorescence using fixed confocal micrographs of UM‐SCC47 cells stained with DAPI, anti‐F‐actin, and anti‐E2F5 antibodies. Images represent at least three independent experiments. Scale bars represent 100 μm.
  3. Protein abundance of E2F5 in cytoplasm versus nucleus in the presence/absence of cisplatin (20 μM, for 0, 1, 2, and 4 h) was detected by Western blotting using cytoplasm‐ versus nuclei‐enriched subcellular fractions of UM‐SCC‐47 cells using anti‐E2F5 antibody. Anti‐clathrin antibody was used to validate cytoplasmic fractions, whereas anti‐lamin B antibody was used to validate nuclear fractions. Western blot images represent at least three independent experiments.