SMN Antibody (2B1) [Alexa Fluor® 647]

Immunocytochemistry/ Immunofluorescence: SMN Antibody (2B1) [Alexa Fluor® 647] [NB100-1936AF647] -

Immunocytochemistry/ Immunofluorescence: SMN Antibody (2B1) [Alexa Fluor® 647] [NB100-1936AF647] - Etv5b activates the smn promoter leading to increased Smn levels in motoneurons.(a–d) Whole-mount in situ hybridization of etv5b expression in 24 hpf embryos. Expression pattern in the entire embryo (a), in magnified lateral (b) & dorsal (c) views as well as in a transverse section (d) of the trunk. In the latter case, yellow arrowhead denote signal in the motoneuron region. Scale bars: 100 μm (a) & 50 μm (b–d). nc, notochord. (e) RT-PCR in control (C - only psmn:mCherry) versus etv5b overexpressing (OE - etv5b mRNA & psmn:mCherry) embryos at early gastrula stage. (f,g) scIF of control (f) & etv5b MO (g) cells from pmnx1:eGFP embryos. H2B, GFP & Smn signals are shown in Z-projected confocal sections. GFP+ cells marked by white rectangles are magnified on the right. Scale bars: 10 μm for low & 5 μm for high magnification. (h) Relative Smn signal in control & in etv5b MO cells from one representative experiment. Exact values are (mean ± SD): 0.99 ± 0.25 (control) & 0.7 ± 0.14 (etvb5 MO), p = 0.003 with Wilcoxon Sum Rank Test. (i) Average relative Smn levels in control & in etv5b MO motoneurons. The exact value of decrease is 0.75 ± 0.04 (mean ± SD). N = 3 experiments, n = number of analyzed cells. Image collected & cropped by CiteAb from the following publication (https://www.nature.com/articles/srep27470), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: SMN Antibody (2B1) [Alexa Fluor® 647] [NB100-1936AF647] -

Immunocytochemistry/ Immunofluorescence: SMN Antibody (2B1) [Alexa Fluor® 647] [NB100-1936AF647] - Motoneurons exhibit elevated Smn levels.(a–c) Single-cell immunofluorescent (scIF) experiments. 24 hpf pmnx1:eGFP embryos were dissociated & immunostained for Smn ((a) see also Methods). DIC image of cells following 3-hour (b) & 24-hour (c) incubation are shown with GFP in green. Note that the GFP signal in the cell body is oversaturated so that the weaker signal in the axon becomes visible. In (c) DNA is in blue & the motoneuron marker Znp1 (synaptotagmin) is in red; the axon growth cone is magnified in the corner of the images. Scale bars are 100 μm (a), 10 μm (low magnification in (b,c)) & 2 μm (high magnification in (c)). (d) scIF on pmnx1:eGFP embryos. Histone 2B (H2B), GFP & Smn signals are shown in Z-projected confocal sections. Cells marked by white rectangles are magnified on the right. Scale bars: 10 μm for low & 5 μm for high magnification. (e) To account for potential variability in the immunostaining, Smn levels were quantified relative to H2B. Diamonds denote GFP negative (GFP−) & GFP positive (GFP+) cells from one representative experiment. Blue bars indicate mean ± SD with significance values of *p < 0.05 & **p < 0.01. Exact values are (mean ± SD) 0.51 ± 0.14 (GFP−) & 0.87 ± 0.17 (GFP+), p = 0.001 with Wilcoxon Sum Rank Test. For more details, see Materials & Methods & Supplementary Table S1. (f) Average increase of Smn levels in motoneurons versus control cells. The exact value of enrichment is (mean ± SD): 1.67 ± 0.14. N = 3 experiments, n = number of analyzed cells. Image collected & cropped by CiteAb from the following publication (https://www.nature.com/articles/srep27470), licensed under a CC-BY license. Not internally tested by Novus Biologicals.