Fig. 8

cPLA2 inhibition blunted the effect of pemetrexed on spheroid formation. Briefly, MSTO-211H cells were clonally seeded in spheroid-forming conditions, in presence or absence of the cPLA2 inhibitor (AVX001 3micromol/L) (upper and middle panels) or in presence of AVX001 + AA (lower panels) and allowed to grow for 7 days, after treatment with ctrl or pem at increasing doses (20–80 micrograms/ml/1 h), before washing out of the drugs. A. Representative micrographs of the formed spheroids at day 5. Scale bar: 200 μm. B. Graphs showing quantitation of the Sphere Forming Efficiency (SFE) from HP1 (left) and MSTO-211H (right) cells seeded and treated as in A (1000 cells/well seeded). Please note that little to no spheroid formation was observed at higher doses of pem, in presence of the cPLA2 inhibitor and that adding back AA rescued cPLA2 inhibition