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Fig. 8. Expression of activated Ras-V12 promotes MEK/ERK-independent survival of 
3-null keratinocytes. 3-null MK-/- cells were infected with adenovirus expressing HA-tagged Ras-V12 (Ras-V12 lanes) or with a control adenovirus expressing ß-galactosidase (ß gal lanes) then sub-cultured on LN-5 ECM for 24 hours in serum-free medium in the presence or absence of 10 µM U0126, as indicated. As controls, uninfected MK-/- cells were cultured in the presence of serum (uninfect., +serum) or in serum-free medium (uninfect., serum-free). (A) To confirm expression of Ras-V12, cell lysates were immunoblotted with anti-HA tag (HA-Ras-V12). ERK activation in Ras V12-infected cells was assayed by probing parallel blots with anti-phospho-ERK (pERK) or total ERK (ERK), as described in Fig. 6. (B) Cells were assayed for activation of caspase-3 by western blot, as described in Fig. 2; migratory positions of procaspase-3 and cleaved caspase-3 are indicated. Filters were stripped and re-probed for keratin 14 (K14). Results are representative of five independent experiments for adenoviral infections; inhibitor treatments were included in two experiments.
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