Table 1.

Summary of Klotho effects on MO3.13 cells

Time	Assay/analysis	Effects	Figure or table
1 h	Western blot, qPCR	Enhancement of Akt and ERK signaling	Fig. 1, Table S1
2.5 h	Microarray, qPCR, Ingenuity pathway analysis, GSEA	80 % of the differentially expressed genes downregulated. High enrichment in the gene set that is also observed in cancer, cardiovascular disease, stress, aging and hormone-related chemical and genetic perturbations	Figs. 2, 3, and 6; S1, Table S1, S3, and S4
3.5 h	qPCR	Similar to 2.5 h	Fig. 3c and Table S1
24 h	Crystal Violet, Luciferase	Cell proliferation not affected. Inhibited: SP1, NFκB, Wnt, VDR, Pax6, Pax3, E2F, p53, AP1 MAPK/JNK, and C/EBP signaling pathways Activated: SRE MAPK/ERK, YY1, PI3K/AKT FOXO, LXR, GATA, and PPAR signaling pathways	Figs. 4 and 5
48 h	Western blot, Crystal Violet, MTT, apoptosis	Decrease in proliferative abilities and increased apoptosis	Fig. 4
1 Week	Western blot	Enhanced differentiation	Fig. 4

GSEA gene set enrichment analysis, crystal Violet staining assay for cell proliferation/cell number, Luciferase luciferase transcription factor pathway reporter assay, MTT cell proliferation/cell viability assay, apoptosis Caspase-glo 3/7 assay