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. 2022 May 26;27(11):3425. doi: 10.3390/molecules27113425

Figure 4.

Figure 4

The effect of 0.3 mM spermidine (SPD) and 0.2 mM eugenol (EUG), alone and in combination (0.3 mM SP + 0.2 mM EUG) on LC3-II activation and viability in Caco-2 cell lines. (A) LC3-II staining in the untreated control cells), and the starvation (STARV)-induced control cells was detected with immunofluorescence green staining (FG) after 4 h. (B,C) For the SPD, EUG and SPD + EUG treatments, LC3-II was detected with FG staining and fast red chromogen staining. (D–F) Statistical analysis of the LC3-II positive cells stained with red chromogen was performed after both 4 h and 24 h from the start of all treatments. MTT assay performed after 24 h (G) and 48 h (H) on Caco-2 cells that were untreated or treated with SPD, EUG and SPD + EUG. The number of stars *, **, *** and **** represents significant differences between treatments as determined by one-way ANOVA at the 99% confidence level (p < 0.01), whereas n.s. is not significant.