Figure 4
From: Regulation of multiple tip formation by caffeine in cellular slime molds
Expression studies. D. discoideum slugs (AX2) were used to perform these assays. A) Western blots: Expression of PdsA protein (AX2) at 0 h, 1 h and 3 h after transfer of slugs to 5 mM caffeine containing plates. B) Real time-PCR: Analysis of AcaA-mRNA, Pde4-mRNA, PkaC-mRNA and 5’NT-mRNA expression levels in multi tipped slugs at 6 h of development. C) AcaA-lacZ staining: Arrow indicates localization of Aca-lacZ in control slugs and the slugs having multiple tips (6 h of development). Reduced AcaA expression in the additional tips. D) cAMP quantification: graph shows absolute concentration of cAMP in the slugs formed from 1X 107 cells. The cAMP levels were quantified in control and multi tipped slugs as mentioned in material and methods. These values represent mean ± standard deviation from 4 independent samples (Student t-test, **P < 0.01).