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Fig. 7. Inhibition of Ins(1,4,5)P₃ and calcium but not PKC inhibits early changes in cell morphology. (A) BE cells plated on Matrigel and treated for 8 hours with either 2-APB (40 µM) an Ins(1,4,5)P₃ receptor antagonist, BAPTA-AM (20 µM), an intracellular calcium-chelating agent, thapsigargin (THAPS; 100 nM) an inhibitor of the sarcoplasmic endoplasmic reticulum calcium ATPase or the PKC inhibitor compounds R0-31-8220 (3 µM) or GF109203X (3 µM) and its structurally related control bisindolylmaleimide V (BiV) (3 µM). Cell elongation was analysed by microscopy with representative images shown and average percentage cell elongation, analysed from 20 independent cell images reported with standard deviations; control 161.2±17.8%, APB-2 7.7±6.4%, BAPTA-AM 1.8±6.5%, thapsigargin 3.2±4.3%, RO-31-8220 45.0±17.9%, GFX109203X 41.0±21.3%, BisV 154±16.7%. (B) HUVECs were plated on Matrigel and treated as for part A. Cell elongation and networking was analysed by fluorescent microscopy. (C) Serum-starved cells treated as for part A were plated onto Matrigel-coated Transwell chambers and invasion through the filter and into the matrix, towards serum (10%) was analysed by microscopy after 48 hours with the average percentage (number) of invading cells compared with control (untreated, 100%) sample from five experiments displayed with standard deviations. 1, control; 2, 2-APB; 3, BAPTA-AM; 4, R0-31-8220; 5, GFX109203X; 6, BiV.

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