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. 2007 Aug 29;8(11):1644–1655. doi: 10.1111/j.1600-0854.2007.00631.x

Figure 8.

Figure 8

Silencing of all three ARFGAPs is lethal in HeLa cells. Silencing of ARFGAP1 or a combination of ARFGAP2 and ARFGAP3 failed to cause lethality in HeLa cells, as well as in NRK cells (data not shown). A) The depletion of ARFGAP1, ARFGAP2 or ARFGAP3 in single knock-downs in HeLa cells was verified by immunoblotting. Antibodies against tubulin and γ-COP were used to verify equal protein loading. B and C) Cell counting was performed from DAPI-stained HeLa cells grown on coverslips. Note the strong reduction in cell numbers in the ARFGAP triple knock-down compared with a double knock-down involving ARFGAP2 and ARFGAP3 combined with a control oligo. For details of the oligos, see Materials and Methods. D) Quantification of the reduction in cell numbers from the above. More than 75% cell death was routinely observed after three sets of siRNA transfections at the 72-h time-point after the last transfection (in 10 independent experiments). Bar = 100 μm.