*4.6. Data Analysis*

The raw data from the microplate reader were exported to Microsoft Excel 2007 (Redmond, WA, USA). From the growth curves, the viability of the yeast can be obtained according to previous report [25]. Survival integral [56] of each aging culture was defined as the area under the survival curves [25,57]. The data were analyzed by one-way analysis of variance (one-way ANOVA), and were expressed as the mean values ± standard error of mean (SEM). The significance of difference (\* *p* < 0.05; \*\* *p* < 0.01; \*\*\* *p* < 0.001; \*\*\*\* *p* < 0.0001) was determined using Sidak's multiple comparisons test. GraphPad Prism 7 (GraphPad Software, Inc., La Jolla, CA, USA) was used for the analysis.

### **5. Conclusions**

In conclusion, neohesperidin, with relatively high capability of removing intracellular ROS, showed great potential in extending the CLS of budding yeast BY4742 individually or synergistically with hesperetin. This might lead to new choices for the treatment of aging problems and since there was a limited relationship between the CLS-extension of yeast and the tested indexes (e.g., ROS scavenging ability, in vitro antioxidant activity and the extracellular pH). Further studies to discover the molecular mechanisms of this phenomenon will be extremely beneficial to prevent aging. Because of this, the importance of choosing the best combination of flavonoids, should be borne in mind when designing new dietary supplements or functional foods.

**Supplementary Materials:** The following are available online at http://www.mdpi.com/1420-3049/24/22/4093/s1, Table S1: Composition of synthetic-defined (SD) medium used for yeast chronological lifespan analysis.

**Author Contributions:** Conceptualization, C.G., H.Z., X.G. and Z.Z.; Data curation, C.G. and X.G.; Formal analysis, C.G. and X.G.; Funding acquisition, H.Z., C.G. and Z.Z.; Methodology, Z.Z., C.G., H.Z. and X.G.; Project administration, Z.Z. and X.G.; Software, X.G.; Supervision, X.G. and Z.Z.; Validation, C.G. and H.Z.; Writing – original draft, Z.Z., X.G. and C.G.; Writing–review & editing, Z.Z., X.G., C.G. and H.Z.

**Funding:** This research was funded by the National Natural Science Foundation (31772260) and Chongqing Graduate Student Innovation Project for Chunxia Guo (CYS16081), the Program for Chongqing Municipal Education Commission (KJQN201901227), and the Foundation of Engineering Technology Research Center of Characteristic Biological Resources in Northeast of Chongqing.

**Acknowledgments:** We appreciated the technical support of Chun Tan in College of Horticulture and Landscape Architecture, Southwest University, China and Charles Wilson in Chongqing Three Gorges University, China.

**Conflicts of Interest:** The authors declare no conflict of interest.
