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. 2021 Jul 23;13(8):2517. doi: 10.3390/nu13082517

Figure 1.

Figure 1

Long-term caffeine intake delays intestinal aging during advanced ages of Caenorhabditis elegans: (A) Animals expressing discs large MAGUK scaffold protein 1 (DLG-1):: green fluorescent protein (GFP) were treated with 0 or 10 mM caffeine at the L4 stage at 25 °C for 72 h. The location of DLG-1::GFP in the pharynx was observed at 72 h post-L4 stage at 25 °C. Error bars represent standard deviation (SD). *** p < 0.001 (one-way analysis of variance (ANOVA) with Tukey’s post hoc test); (B) The synchronized wild-type L4-stage animals were treated with 0 or 10 mM caffeine at 25 °C for 72 h. The intestinal atrophy was measured at 72 h post-L4 stage at 25 °C. Error bars represent SD. *** p < 0.001. n.s., not significant (two-way ANOVA with Tukey’s post hoc test); (C) Animals expressing actin 5 (ACT-5)::GFP were treated with 0 or 10 mM caffeine at the L4 stage at 25 °C for 72 h. The type of mislocalization was classified into four stages. The percent distributions of the respective stages in animals fed with 0 or 10 mM caffeine are presented; (D) The synchronized wild-type L4-stage animals were fed with E. coli OP50::GFP, a fluorescent bacteria on 0 or 10 mM caffeine nematode growth medium (NGM) plates at 25 °C for 72 h. The type of bacterial colonization was classified into three categories: (1) undetectable, (2) partial, and (3) full. The percent distributions of the respective categories in animals treated with 0 or 10 mM caffeine are presented; (E) The synchronized wild-type L4-stage animals were treated with 0 or 10 mM caffeine at 25 °C for 72 h. Accumulation of the pseudocoelomic lipoprotein pool (PLP) (indicated by yellow arrowheads) was observed at 72 h in post-L4-stage animals at 25 °C. The percentage of animals with PLP accumulation among the total number of animals is shown. Error bars represent SD. *** p < 0.001 (t-test).