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. 2020 Jan 13;12(2):e10812. doi: 10.15252/emmm.201910812

Figure EV5. (supporting Fig 6). Combined depletion of Vps4a+b in mouse colon carcinoma CT‐26 cells.

Figure EV5

  • A
    qRT–PCR analysis of the silencing efficiency of Vps4a (upper panel) and Vps4b (lower panel) in CT‐26 cells 72 h after transfection with siRNA. To deplete Vps4a or Vps4b, two independent siRNA duplexes were used (#2 or #5, and #3 or #4, respectively). To simultaneously deplete Vps4a+b, various combinations of siVps4a and siVps4b were used. All values were normalized; Vps4a or Vps4b expression values in siCTRL#1‐transfected cells were set as 1 and used to normalize mRNA abundance in other conditions. NT—non‐transfected.
  • B
    Immunoblotting detection of Vps4a abundance, inflammatory response (phosphorylated RelA) and apoptosis activation (cleaved caspase 7) in lysates of mouse CT‐26 cells collected 72 h after siRNA transfection as in (A). p‐RelA—phosphorylated RelA; cl—cleaved caspase. Vinculin served as a loading control.
  • C
    Phase contrast microscopy images of CT‐26 cells acquired 3 days after transfection with siVps4a or siVps4b as described in (A). Scale bar, 250 μm.