Fig. 5: Differential processing of dead neurons in ALS-PFN1 mouse brains compared to controls.
From: Expression of ALS-PFN1 impairs vesicular degradation in iPSC-derived microglia
Analysis of PFN1 C71G+/− (n = 4) and WT (n = 5) mouse brains injected with dead neurons co-labeled with the constitutively fluorescent dye Alexa Fluor 546 (AF-neurons) and the pH-sensitive dye Cypher5E (Cy5E-neurons) 72 h post-injection. a Representative immunofluorescence images at the injection site in the mouse motor cortex of the myeloid marker IBA1(gray), the residual dead neurons (AF-neurons in green), and the dead neurons localized in acidic compartments (Cy5E-neurons in red) as well as the overlaid image of AF-neurons and Cy5E-neurons (merged in yellow). Scale bar: 200 µm. b Representative confocal images of IBA1-positive (IBA1+) cells colocalizing with Cy5E-neuron signal from a PFN1 WT (top) and PFN1 C71G+/− (bottom) mouse brain (scale bar: 25 µm) including higher magnification insets (scale bar: 10 µm). c Quantification of the total area of residual AF-neuron signal (ns P = 0.7288, t = 0.3610, df = 7). d The fraction of Cy5E-neuron (total area) signal that overlays with the AF-neuron (total area) signal from c (*P = 0.0499, t = 2.366, df = 7). e Representative IBA1 (gray) immunofluorescence images at the injection site for WT PFN1 and PFN1 C71G+/− mice. Three concentric rings (yellow) around the site of injection (center ring) are shown for analysis in (f and g). Scale bar: 100 µm. f Quantification of total IBA1 signal intensity in the three rings surrounding the injection site labeled in (e) normalized to the number of IBA+ cells (**P = 0.0058, t = 3.909, df = 7 for ring 1; *P = 0.0264, t = 2.804, df = 7 for ring 2; *P = 0.0374, t = 2.563, df = 7 for ring 3). g Quantification of the number of IBA1+ cells in each ring indicated in (e) (ns P = 0.4752, t = 0.7545, df = 7 for ring 1; ns P = 0.9264, t = 0.09573, df = 7 for ring 2; *P = 0.0255, t = 2.827, df = 7). Unpaired two-tailed t-test was used for all statistical comparisons. Graphs in this figure show mean ± SEM. Data points represent individual animals. Males are plotted open symbols and females with closed symbols. Source data are provided as a Source Data file.
