Figure 1

Effect of caffeine on multiple tip formation in D. discoideum. A) Monitoring multiple tip formation in D.discoideum. Freshly harvested cells were stained with neutral red (0.06%) and allowed to form slugs. For getting multiple tip effect, fully developed slugs were transferred with a fine needle onto a non-nutrient agar plate containing 5 mM caffeine as described in the material and methods. Multiple tip formation was monitored by taking pictures at the indicated time points in development. Arrows in Figure 1A represent the emergence of secondary tips at respective stage of development at the mentioned time course. Scale bar = 200 μm. B) Graph shows the number of multiple tips in slugs at different concentrations of caffeine at indicated time points. At each caffeine concentration, 15 slugs were transferred for secondary tip formation and were monitored at 3 h, 4 h, 5 h, and 6 h of development. These values represent mean ± standard deviation.