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Detection of Mouse SOX10 by Immunohistochemistry

PDGFR alpha driven brain tumors display features of high grade glioma.(a–g) Histopathological analysis of tumor areas by H&E staining shows a high concentration of mitotic figures (a, arrows), high cellularity and nuclear atypia (b), perineuronal satellitosis (c; N, neuronal nuclei), perivascular growth (d), intrafascicular growth (e), subarachnoid spreading (f), and areas of incipient necrosis (g; arrows point to pyknotic nuclei). (h–k) IF labeling of brain tumor sections for cell type specific markers. Nuclei labeled with DAPI are shown in blue. Tumor cells with high PDGFR alpha expression were highly proliferative, as seen by proliferation marker Ki67 (h), and express the OPC cell lineage markers Olig2, Sox2, Sox10, and Ng2, as well as the neural stem cell marker Nestin (i–k). Tumor cells were negative for immunosignal of astroglial marker GFAP, mature oligodendrocyte marker APC-CC1, and neuronal marker NeuN (l–n). Scale bars: 10 μm (a–g), 20 μm (h–n). Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/25683249), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse SOX10 by Immunocytochemistry/Immunofluorescence

Adult dynamin 2-depleted Schwann cells are replaced by dynamin 2-positive Schwann cells in remyelinated nerves.(A) Immunostainings for DNM2, SOX10 (SCs) and YFP (reporter-recombined SCs) on SN cross-sections of control* (MpzCreERT2:Rosa26-stoploxP/loxP-YFP) and P0ERT2-Dnm2KO* (MpzCreERT2:Dnm2loxP/loxP: Rosa26-stoploxP/loxP-YFP) mice at 4 wpT and 14 wpT. White arrows: YFP-recombined SCs that express DNM2 (in controls at 4 wpt and 14 wpt); black arrowheads: YFP-recombined SCs lacking DNM2 (in mutants at 4 wpt); white arrowheads: Non-recombined SCs expressing DNM2 (in controls and mutants at 4 wpt and 14 wpt). Scale bar = 25 μm for entire panel. (B) Quantification related to (A). Percentage of YFP+ cells among SOX10+ SCs/SN cross-sections at 4 wpT, 6 wpT, and 14 wpT. N = 3 mice/genotype, Two-Way ANOVA with Sidak’s multiple comparisons test. (C) Western blot analysis of cleaved caspase 3 (cC3) in SN lysates of control* and P0ERT2-Dnm2KO* mice at 4 wpT, 6 wpT and 14 wpT. Full-length blots in Supplementary file 1F. (D) Quantification referring to (C). Control average was set to 1. N = 3 mice/time point and genotype, Two-Way ANOVA with Sidak’s multiple comparisons test. Results in graphs represent means ±s.e.m.; ***p<0.001. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30648534), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse SOX10 by Immunocytochemistry/Immunofluorescence

Schwann cells lacking dynamin 2 show impaired cell cycle progression, reduced mitosis rate, and cytokinesis defects.(A) Schematic representation of the cell cycle phases marked by Ki-67 and EdU. (B) EdU-labeling, combined with immunostainings for Ki-67 and SOX10 on control and P0-Dnm2KO SN cross-sections at P5. Arrows: EdU+ Ki-67+ SOX10+ SCs, arrowheads: Ki-67+ SOX10+ SCs. Scale bar = 25 μm for entire panel. (C) Quantification of (B). Percentage of EdU+ SCs/SN cross-sections. N = 5 mice/time point and genotype, Two-Way ANOVA with Sidak’s multiple comparisons test. (D) Quantification of (B). Percentage of Ki-67+ SCs/SN cross-sections at P5. N = 5 mice/genotype, two-tailed unpaired Student´s t-test. (E) Quantification of (B). Percentage of EdU+ among Ki-67+ SCs/SN cross-section at P5. N = 5 mice/genotype, two-tailed unpaired Student´s t-test. (F) Quantification of mitotic events in cultured mouse SCs isolated from MpzCre:Rosa26-stoploxP/loxP-YFP (control*) and MpzCre:Dnm2loxP/loxP:Rosa26-stoploxP/loxP-YFP (P0-Dnm2KO*) SNs at P1, monitored by time-lapse microscopy for 24 hr. Each data point represents one individual animal (at least 42 cells/animal analyzed). N = 5 controls and seven mutant mice, two-tailed unpaired Student´s t-test. (G) Quantification of mitosis duration (minutes) of SCs of control* and P0-Dnm2KO* mice, monitored by time-lapse microscopy for 24 hr. Each data point represents one cell derived from a total of 5 control and seven mutant mice. Cells derived from each animal were isolated and analyzed separately, but pooled in one graph; two-tailed unpaired Student´s t-test. (H) Exemplary picture of a multinucleated Dnm2KO SCs after 48 hr in culture. Scale bar = 25 μm. (I) Quantification of multinucleated cells in control and Dnm2KO SCs after 24 hr, 48 hr and 76 hr in culture. N = 6 mice/genotype for 48 hr; N = 6 control* and n = 5 P0-Dnm2KO* mice for 24 hr and 72 hr, two-Way ANOVA with Sidak’s multiple comparisons test. (J) Exemplary time-lapse images of the mitosis of control and Dnm2KO SCs (YFP+ cells). Arrowheads: cell body of dividing SCs, black arrows: cytokinesis site. (K) Representative time-lapse images of Dnm2KO SC (YFP+) failing cytokinesis. The SC is undergoing mitosis, but fails to divide (arrowheads point to the cell body). Results in graphs represent means ±s.e.m.; *p<0.05, ***p<0.001. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30648534), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse SOX10 by Immunohistochemistry

PDGFR alpha driven brain tumors display features of high grade glioma.(a–g) Histopathological analysis of tumor areas by H&E staining shows a high concentration of mitotic figures (a, arrows), high cellularity and nuclear atypia (b), perineuronal satellitosis (c; N, neuronal nuclei), perivascular growth (d), intrafascicular growth (e), subarachnoid spreading (f), and areas of incipient necrosis (g; arrows point to pyknotic nuclei). (h–k) IF labeling of brain tumor sections for cell type specific markers. Nuclei labeled with DAPI are shown in blue. Tumor cells with high PDGFR alpha expression were highly proliferative, as seen by proliferation marker Ki67 (h), and express the OPC cell lineage markers Olig2, Sox2, Sox10, and Ng2, as well as the neural stem cell marker Nestin (i–k). Tumor cells were negative for immunosignal of astroglial marker GFAP, mature oligodendrocyte marker APC-CC1, and neuronal marker NeuN (l–n). Scale bars: 10 μm (a–g), 20 μm (h–n). Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/25683249), licensed under a CC-BY license. Not internally tested by R&D Systems.