Research Paper Volume 13, Issue 10 pp 13474—13495

Ganoderma lucidum stimulates autophagy-dependent longevity pathways in Caenorhabditis elegans and human cells

G. lucidum treatment reduces FIB-1 levels and extends lifespan in C. elegans. (A) Effects of a water extract of G. lucidum (GL) on fibrillarin-1 (FIB-1) levels as monitored by fluorescence microscopy. Synchronized L4 larvae of transgenic C. elegans strain SJL1 expressing FIB-1::GFP (green fluorescent protein) under the FIB-1 gene’s native promoter were cultured for 3 days on agar plates spread with control water, the water extract of GL, or rapamycin (400 μM, Rapa). Data are expressed as arbitrary units (a. u.). (B) FIB-1 levels assessed by Western blots in wild-type N2 C. elegans treated with GL. (C) Quantification of FIB-1 protein levels shown in (B). FIB-1 expression was measured by densitometry and normalized against actin. (D) Lifespan assay of GL-treated nematodes. C. elegans was cultured on agar plates spread with water, GL or rapamycin as above. Survival was assessed for 30 days using an optical microscope based on motility. Representative lifespan curves are shown. See also Supplementary Table 1. (E) Pharyngeal pumping of nematodes following culture with water, GL or rapamycin for 3 days. Pharyngeal contractions were recorded for 1 min under optical microscopy. (F) Size of 3-day old worms. Size was monitored by delineating the worms’ region of interest (ROI) under optical microscopy. Data represent means ± standard deviation. *p

Figure 1. G. lucidum treatment reduces FIB-1 levels and extends lifespan in C. elegans. (A) Effects of a water extract of G. lucidum (GL) on fibrillarin-1 (FIB-1) levels as monitored by fluorescence microscopy. Synchronized L4 larvae of transgenic C. elegans strain SJL1 expressing FIB-1::GFP (green fluorescent protein) under the FIB-1 gene’s native promoter were cultured for 3 days on agar plates spread with control water, the water extract of GL, or rapamycin (400 μM, Rapa). Data are expressed as arbitrary units (a. u.). (B) FIB-1 levels assessed by Western blots in wild-type N2 C. elegans treated with GL. (C) Quantification of FIB-1 protein levels shown in (B). FIB-1 expression was measured by densitometry and normalized against actin. (D) Lifespan assay of GL-treated nematodes. C. elegans was cultured on agar plates spread with water, GL or rapamycin as above. Survival was assessed for 30 days using an optical microscope based on motility. Representative lifespan curves are shown. See also Supplementary Table 1. (E) Pharyngeal pumping of nematodes following culture with water, GL or rapamycin for 3 days. Pharyngeal contractions were recorded for 1 min under optical microscopy. (F) Size of 3-day old worms. Size was monitored by delineating the worms’ region of interest (ROI) under optical microscopy. Data represent means ± standard deviation. *p<0.05; **p<0.01; ***p<0.001.