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a , qRT-PCR analyses of pro-autophagic genes in old HSCs (n = 3–5). Results are expressed as log2 fold expression compared to young HSCs (set to 0). b , Apoptosis levels in cultured young and old HSCs (n = 3). Results are expressed as percent caspase activation compared to +cytokines young HSCs. Hatching indicates –cytokines conditions. c , 2-NBD glucose uptake in cultured young and old HSCs (n = 3). d , Percent colony formation in young and old WT HSCs cultured for 32h ± BafA and <t>methylpyruvate</t> (M-Pyr) (n = 3). Colonies were counted at day 10 and normalized to -BafA conditions. Data are means ± s.d. *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001.
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a , qRT-PCR analyses of pro-autophagic genes in old HSCs (n = 3–5). Results are expressed as log2 fold expression compared to young HSCs (set to 0). b , Apoptosis levels in cultured young and old HSCs (n = 3). Results are expressed as percent caspase activation compared to +cytokines young HSCs. Hatching indicates –cytokines conditions. c , 2-NBD glucose uptake in cultured young and old HSCs (n = 3). d , Percent colony formation in young and old WT HSCs cultured for 32h ± BafA and <t>methylpyruvate</t> (M-Pyr) (n = 3). Colonies were counted at day 10 and normalized to -BafA conditions. Data are means ± s.d. *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001.
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a , qRT-PCR analyses of pro-autophagic genes in old HSCs (n = 3–5). Results are expressed as log2 fold expression compared to young HSCs (set to 0). b , Apoptosis levels in cultured young and old HSCs (n = 3). Results are expressed as percent caspase activation compared to +cytokines young HSCs. Hatching indicates –cytokines conditions. c , 2-NBD glucose uptake in cultured young and old HSCs (n = 3). d , Percent colony formation in young and old WT HSCs cultured for 32h ± BafA and methylpyruvate (M-Pyr) (n = 3). Colonies were counted at day 10 and normalized to -BafA conditions. Data are means ± s.d. *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001.

Journal: Nature

Article Title: FoxO3a Directs a Protective Autophagy Program in Hematopoietic Stem Cells

doi: 10.1038/nature11895

Figure Lengend Snippet: a , qRT-PCR analyses of pro-autophagic genes in old HSCs (n = 3–5). Results are expressed as log2 fold expression compared to young HSCs (set to 0). b , Apoptosis levels in cultured young and old HSCs (n = 3). Results are expressed as percent caspase activation compared to +cytokines young HSCs. Hatching indicates –cytokines conditions. c , 2-NBD glucose uptake in cultured young and old HSCs (n = 3). d , Percent colony formation in young and old WT HSCs cultured for 32h ± BafA and methylpyruvate (M-Pyr) (n = 3). Colonies were counted at day 10 and normalized to -BafA conditions. Data are means ± s.d. *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001.

Article Snippet: Bafilomycin A (BafA, 5 nM, Sigma), methylpyruvate (M-Pyr, 10 mM, Sigma), rapamycin (Rapa, 20 μM, EMD Millipore) or appropriate vehicles (DMSO for BafA and Rapa) were added to the media as indicated.

Techniques: Quantitative RT-PCR, Expressing, Cell Culture, Activation Assay