Journal: Acta Neuropathologica Communications

Article Title: Absence of Ledgf in mouse brain affects the Kmt2a/b and polycomb balance, synaptic transmission and motor function

doi: 10.1186/s40478-025-02216-4

Figure Lengend Snippet: Validation of Psip1 c KO in the central nervous system. A Western Blot for LEDGF and vinculin expression in different brain regions from 12 week old animals. n = 4 for each brain region to calculate the ratio of the isoforms. B Western Blot for LEDGF and vinculin expression in spinal cord from 8 week old animals. C Validation of genotype on genomic DNA. Top panel: PCR for Psip1 , resulting in a band of 745 bp for Psip1 WT/WT and a band of 802 bp for Psip1 fl/fl mice. Bottom panel: PCR for Cre, resulting in a band of 350 bp for Nestin Cre/− mice. D Staining of different brain regions and spinal cord for LEDGF/p75 specifically or LEDGF/p75 & LEDGF/p52 in WT and cKO animals at an age of 8 weeks. NeuN was used as a neuronal marker. Merged images highlight the co-localization of LEDGF and NeuN. CB = cerebellum; HIP = hippocampus; STR = striatum; CTX = cortex and OB = olfactory bulb. Scale bar is 100 µm. Images are taken with a fluorescence microscope with the 5X objective.

Article Snippet: To assess the expression of both isoforms the rabbit anti-LEDGF/p75 antibody (Bethyl, A300-848A) was used in a dilution of 1/500 and the mouse anti-Psip1 antibody (Invitrogen, MA5-15,795) was used in a dilution of 1/1000.

Techniques: Biomarker Discovery, Western Blot, Expressing, Staining, Marker, Olfactory, Fluorescence, Microscopy

Journal: Acta Neuropathologica Communications

Article Title: Absence of Ledgf in mouse brain affects the Kmt2a/b and polycomb balance, synaptic transmission and motor function

doi: 10.1186/s40478-025-02216-4

Figure Lengend Snippet: Validation of Psip1 c KO in the central nervous system. A Western Blot for LEDGF and vinculin expression in different brain regions from 12 week old animals. n = 4 for each brain region to calculate the ratio of the isoforms. B Western Blot for LEDGF and vinculin expression in spinal cord from 8 week old animals. C Validation of genotype on genomic DNA. Top panel: PCR for Psip1 , resulting in a band of 745 bp for Psip1 WT/WT and a band of 802 bp for Psip1 fl/fl mice. Bottom panel: PCR for Cre, resulting in a band of 350 bp for Nestin Cre/− mice. D Staining of different brain regions and spinal cord for LEDGF/p75 specifically or LEDGF/p75 & LEDGF/p52 in WT and cKO animals at an age of 8 weeks. NeuN was used as a neuronal marker. Merged images highlight the co-localization of LEDGF and NeuN. CB = cerebellum; HIP = hippocampus; STR = striatum; CTX = cortex and OB = olfactory bulb. Scale bar is 100 µm. Images are taken with a fluorescence microscope with the 5X objective.

Article Snippet: To assess the expression of both isoforms the rabbit anti-LEDGF/p75 antibody (Bethyl, A300-848A) was used in a dilution of 1/500 and the mouse anti-Psip1 antibody (Invitrogen, MA5-15,795) was used in a dilution of 1/1000.

Techniques: Biomarker Discovery, Western Blot, Expressing, Staining, Marker, Olfactory, Fluorescence, Microscopy

Journal: Acta Neuropathologica Communications

Article Title: Absence of Ledgf in mouse brain affects the Kmt2a/b and polycomb balance, synaptic transmission and motor function

doi: 10.1186/s40478-025-02216-4

Figure Lengend Snippet: Expression of LEDGF/p75 in specific neuronal subtypes. A Co-staining for choline acetyltransferase (ChAT) and LEDGF/p75 in motor neurons of the spinal cord of WT and cKO mice. B Co-staining for calbindin and LEDGF/p75 in the Purkinje cell layer of the cerebellum of WT and cKO mice. ML = granular layer; PL = Purkinje cell layer; GL = granular layer C Co-staining for tyrosine hydroxylase (TH) and LEDGF/p75 in dopaminergic neurons in the substantia nigra pars compacta in WT and cKO mice. D Example of TH staining in the substantia nigra of WT and cKO mice of 1 year of age. E Number of TH positive neurons in the right hemisphere of WT and cKO mice at 1 year of age. Unpaired t-test with n = 8 for WT/n = 9 for cKO. Data are presented as mean ± SEM and individual values. Scale bar is 50 µm. Images taken with confocal microscope with 20X objective.

Article Snippet: To assess the expression of both isoforms the rabbit anti-LEDGF/p75 antibody (Bethyl, A300-848A) was used in a dilution of 1/500 and the mouse anti-Psip1 antibody (Invitrogen, MA5-15,795) was used in a dilution of 1/1000.

Techniques: Expressing, Staining, Microscopy

Journal: Acta Neuropathologica Communications

Article Title: Absence of Ledgf in mouse brain affects the Kmt2a/b and polycomb balance, synaptic transmission and motor function

doi: 10.1186/s40478-025-02216-4

Figure Lengend Snippet: Model for LEDGF mediated control of gene expression. In WT mice transcription is regulated by keeping the trithorax and polycomb repressor complex 2 (PRC2) in balance. LEDGF/p75 act as a molecular tether to guide the trithorax system to active genes. However, in Psip1 cKO mice, LEDGF/p75 is no longer present resulting in the loss of binding of the trithorax system to chromatin. As a result, the PRC2 complex is more active resulting in more downregulated genes. Ezh2 = enhancer of zeste homolog 2, Eed = embryonic ectoderm development, Suz12 = suppressor of zeste 12, RbAp46 = Histone binding protein, SET = Sur3-9 Enhancer-of-zeste, Ash2l = ASH2 Like, Rbbp5 = Retinoblastoma-Binding Protein 5, Dpy30 = dpy-30 histone methyltransferase complex regulatory subunit, Wdr5 = WD Repeat Domain 5.

Article Snippet: To assess the expression of both isoforms the rabbit anti-LEDGF/p75 antibody (Bethyl, A300-848A) was used in a dilution of 1/500 and the mouse anti-Psip1 antibody (Invitrogen, MA5-15,795) was used in a dilution of 1/1000.

Techniques: Control, Gene Expression, Binding Assay