Fig. 4: GDF11 stimulation of neuronal activity is mediated by autophagy. | Nature Aging

Fig. 4: GDF11 stimulation of neuronal activity is mediated by autophagy.

From: Systemic GDF11 attenuates depression-like phenotype in aged mice via stimulation of neuronal autophagy

Fig. 4

a, Representative confocal images of GFP+ dendrites and spines from hippocampal neurons in culture transfected with either shBeclin 1 or GFP (green) and treated with either GDF11 or control (vehicle). Scale bar: 4 μm. b, Quantification of dendritic spine density after a 2-h stimulation with either GDF11 or control (number represents number of spines for every 10 μm of dendrite; nCtrl = 31, nshBec = 18, nGDF11 = 36, nshBec+GDF11 = 35 neurons examined over three independent experiments; F (3, 116) = 8.2). c, Representative confocal images of hippocampal neurons in vitro treated with vehicle, Baf or GDF11 or both and immunostained with cFos (green) and Hoechst (blue). Scale bar: 100 μm. d, Quantification of % cFos+ neurons per field (nCtrl = 31 fields, nBaf = 28 fields, nGDF11 = 28 fields, nBaf+GDF11 = 32 fields; F (3, 115) = 7.2). e, Oxygen consumption rate (OCR) in primary hippocampal neurons in vitro treated for 2 h with either GDF11 or Baf or both and measured with the Seahorse analyzer (nCtrl = 20, nBaf = 8, nGDF11 = 8, nBaf+GDF11 = 8 biologically independent samples). f, Measurement of basal OCR before oligomycin/FCCP addition (for each condition the three measurements were pooled; nCtrl = 24, nBaf = 21, nGDF11 = 21, nBaf+GDF11 = 24 biologically independent samples; F (3, 86) = 5.5). g, GSEA from RNA-seq on neurons treated with GDF11 or vehicle for 2 h. h, Western blots images of lysates from hippocampal neurons treated with GDF11 or control (vehicle). i–k, Quantification of western blots by optical density for Deptor (i) (n = 4 biologically independent samples), phospho-S6K1 (j) (n = 8 biologically independent samples) and 4E-BP1 (k) (n = 4 biologically independent samples). Baf refers to Baf. One-way one-sided ANOVA and Tukey’s post hoc test for multiple comparisons; F (DFn, DFd) values presented for each ANOVA statistical analysis; two-sided Mann–Whitney test for two-group comparisons; P values < 0.05 are represented on the graph; mean ± s.e.m.

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