Fig 1: S100A6 signaling is modulated by eicosapentaenoic acid.a Neuritogenesis in cortical neurons (prepared at E18.5) treated with EPA at either 1 μM or 5 μM concentration, and monitored by dynamic live-cell imaging for 7 days in vitro (DIV). Data were normalized to neurite length prior to drug exposure on 1 DIV. Neurite outgrowth was determined by using a manual overlay for each neuron (n = 7 independent experiments). b Cultured mouse astrocytes were stimulated with EPA (30 μM) vs. control. Data were normalized to cell density prior to EPA exposure on 1 DIV (n = 8 independent experiments). c ELISA of S100A6 in culture media upon EPA (30 μM) applied for 24 h (n = 3 biological replicates). d mRNA levels for Cacybp in cortical neurons after exposure to a medium pre-conditioned for 24 h (n = 3 biological replicates). e mRNA levels for Grp78, Atf4, Atf6a, Chop (p = 0.067), Xbp1u, and Xbp1s (p = 0.122) in E18.5 cortical neurons after exposure to astrocyte-conditioned medium for 24 h (n = 3 biological replicates). Mixed-sex embryos were used where relevant. Data were expressed as means ± s.d. throughout. Data were statistically evaluated using ANOVA (a–c) or two-tailed Student’s t-test (d, e), with only post-hoc comparisons indicated (/p < 0.05, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. Unprocessed data and statistics were included in the Source Data File.
Fig 2: S100A6 availability modulates neuronal differentiation.a Co-culture experiments schema. b S100A6 (10 kDa) WB in astrocyte cytosol after 96 h exposure to scrambled or targeting siRNA, followed by 6 h EPA (30 μM) treatment. Representative blots are shown; lane numbers correspond to subsequent panels for clarity. c S100A6 ELISA in culture media upon treatment as in (b) with either n = 4 biological replicates [control] or n = 3 biological replicates [siRNA; EPA; EPA + siRNA]. d S100A6 detection in the cytosol of astrocytes after treatments as in (b) (n = 3 biological replicates). e Representative imagines of cortical neurons exposed to conditioned media for 24 h or EPA (30 μM), and double-labeled for F-actin (phalloidin; magenta) and MAP2 (green). Lead neurites and growth cones were marked by dashed lines and solid arrowheads, respectively. Scale bars = 12 μm (top), 5 μm (bottom). f Primary neurite length after experimental manipulations (biological replicates: n = 7 [control]; n = 5 [cond. siRNA; cond. siRNA + EPA; EPA]; n = 9 [cond. EPA]). g Neuritogenesis after genetic and pharmacological manipulations and monitored for 24 h. Data were normalized to neurite length prior to treatment and expressed in percentage. Neurite outgrowth was determined by using a manual overlay (biological replicates: n = 7 [control; cond. siRNA], n = 4 [cond. EPA], n = 5 [cond. siRNA + EPA], n = 8 [EPA]). Data were expressed as means ± s.d. throughout. Samples were mixed for sex where relevant. Data were statistically evaluated by one-way ANOVA followed by Tukey’s multiple comparisons with post-hoc comparisons indicated (*p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. Unprocessed data and detailed statistics were included in the Source Data File.
Supplier Page from Abcam for Recombinant Mouse S100 alpha 6/PRA protein (His tag N-Terminus)