Journal: Neuron

Article Title: Rewiring of human neurodevelopmental gene regulatory programs by Human Accelerated Regions (HARs)

doi: 10.1016/j.neuron.2021.08.005

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: CaptureMPRA library design, cloning, and transfection Molecular Inversion Probes (MIPs) targeting flanking regions of all HAR sequences were designed using the MIPgen program ( Boyle et al., 2014 ) and manufactured as a pool by CustomArray (Bothell, WA).

Techniques: Virus, Recombinant, Electron Microscopy, Modification, Lysis, Sonication, Plasmid Preparation, Software

Journal: Neuron

Article Title: Rewiring of human neurodevelopmental gene regulatory programs by Human Accelerated Regions (HARs)

doi: 10.1016/j.neuron.2021.08.005

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: CaptureMPRA library design, cloning, and transfection Molecular Inversion Probes (MIPs) targeting flanking regions of all HAR sequences were designed using the MIPgen program ( Boyle et al., 2014 ) and manufactured as a pool by CustomArray (Bothell, WA).

Techniques: Virus, Recombinant, Electron Microscopy, Modification, Lysis, Sonication, Plasmid Preparation, Software

Journal: Histochemistry and Cell Biology

Article Title: Involvement of membrane palmitoylated protein 6 (MPP6) in synapses of mouse cerebrum

doi: 10.1007/s00418-025-02378-1

Figure Lengend Snippet: Western blot analysis of MPP6 in the cerebrum of Mpp6 +/+ and Mpp6 −/− mice. Four primary anti-MPP6 antibodies ( no. 1 (lanes 1, 2), no. 2 (lanes 3, 4), no. 3 (lanes 5, 6), and no. 4 (lanes 7, 8) ) targeting different domains of the MPP6 protein were used. Detailed antibody information is provided in the Materials and methods section

Article Snippet: The membranes were probed with the following primary antibodies (Abs): rabbit polyclonal anti-MPP6 Abs targeting different regions [N-terminal region (MPP6 Ab no. 1: GeneTex; no. GTX108010), second L27 domain (MPP6 Ab no. 2: ATLAS (Stockholm, Sweden); no. HPA019085), central region (MPP6 Ab no. 3: GeneTex; no. GTX117969), C-terminal region (MPP6 Ab no. 4: originally produced by Saitoh et al. ( )], rabbit polyclonal anti-MPP1 Ab (originally produced by Zhang et al. , rabbit polyclonal anti-MPP2 Ab [GeneTex; no. GTX103908 (Yamada et al. )], rabbit polyclonal anti-Lin7 Ab (GeneTex; no. GTX117114), mouse monoclonal anti-CASK Ab (NeuroMab, Davis, CA, USA; no. K56A/50), mouse monoclonal anti-actin Ab (Wako, Osaka, Japan; clone no. 2F3).

Techniques: Western Blot

Journal: Histochemistry and Cell Biology

Article Title: Involvement of membrane palmitoylated protein 6 (MPP6) in synapses of mouse cerebrum

doi: 10.1007/s00418-025-02378-1

Figure Lengend Snippet: Western blot analysis of MPP family proteins in the synaptic fraction. MPP6 ( lanes 1–3 ), MPP1 ( lanes 4–6 ), MPP2 ( lanes 7–9 ), and CASK ( lanes 10–12 ) were analyzed in mouse cerebral lysate ( C; lanes 1, 4, 7, and 10 ) and in the synaptic plasma membrane fraction from the mouse ( mP; lanes 2, 5, 8, and 11 ) and rat ( rP; lanes 3, 6, 9, and 12 ) cerebrum. Asterisks on the right side of lanes 3, 6, 9, and 12 indicate the predicted molecular weights of the MPP6, MPP1, MPP2, and CASK proteins, respectively

Article Snippet: The membranes were probed with the following primary antibodies (Abs): rabbit polyclonal anti-MPP6 Abs targeting different regions [N-terminal region (MPP6 Ab no. 1: GeneTex; no. GTX108010), second L27 domain (MPP6 Ab no. 2: ATLAS (Stockholm, Sweden); no. HPA019085), central region (MPP6 Ab no. 3: GeneTex; no. GTX117969), C-terminal region (MPP6 Ab no. 4: originally produced by Saitoh et al. ( )], rabbit polyclonal anti-MPP1 Ab (originally produced by Zhang et al. , rabbit polyclonal anti-MPP2 Ab [GeneTex; no. GTX103908 (Yamada et al. )], rabbit polyclonal anti-Lin7 Ab (GeneTex; no. GTX117114), mouse monoclonal anti-CASK Ab (NeuroMab, Davis, CA, USA; no. K56A/50), mouse monoclonal anti-actin Ab (Wako, Osaka, Japan; clone no. 2F3).

Techniques: Western Blot, Clinical Proteomics, Membrane

Journal: Histochemistry and Cell Biology

Article Title: Involvement of membrane palmitoylated protein 6 (MPP6) in synapses of mouse cerebrum

doi: 10.1007/s00418-025-02378-1

Figure Lengend Snippet: Immunoprecipitation analysis of MPP6 in the cerebrum. Western blot detection using anti-MPP1 ( lanes 1–4 ), MPP2 ( lanes 5–8 ), CASK ( lanes 9–12 ), and no primary antibody ( lanes 13–15 ) following immunoprecipitation of cerebral lysates with anti-MPP6 antibody. Sample A ( lanes 2, 6, 10, and 13 ): cerebral lysate immunoprecipitated with anti-MPP6 antibody, followed by protein G-sepharose collection. Sample B ( lanes 3, 7, 11, and 14 ): cerebral lysate processed without anti-MPP6 antibody, followed by protein G-sepharose collection. Sample C ( lanes 4, 8, 12, and 15 ): anti-MPP6 antibody alone, collected with protein G-sepharose. Sample L ( lanes 1, 5, and 9 ): cerebral lysate. M : molecular marker. Asterisks on the right side of lanes 2, 6, and 10 indicate bands corresponding to MPP1, MPP2, and CASK. The exposure time for chemiluminescence detection was extended to ensure the detection of faint signals, leading to red-saturated areas

Article Snippet: The membranes were probed with the following primary antibodies (Abs): rabbit polyclonal anti-MPP6 Abs targeting different regions [N-terminal region (MPP6 Ab no. 1: GeneTex; no. GTX108010), second L27 domain (MPP6 Ab no. 2: ATLAS (Stockholm, Sweden); no. HPA019085), central region (MPP6 Ab no. 3: GeneTex; no. GTX117969), C-terminal region (MPP6 Ab no. 4: originally produced by Saitoh et al. ( )], rabbit polyclonal anti-MPP1 Ab (originally produced by Zhang et al. , rabbit polyclonal anti-MPP2 Ab [GeneTex; no. GTX103908 (Yamada et al. )], rabbit polyclonal anti-Lin7 Ab (GeneTex; no. GTX117114), mouse monoclonal anti-CASK Ab (NeuroMab, Davis, CA, USA; no. K56A/50), mouse monoclonal anti-actin Ab (Wako, Osaka, Japan; clone no. 2F3).

Techniques: Immunoprecipitation, Western Blot, Marker

Journal: Histochemistry and Cell Biology

Article Title: Involvement of membrane palmitoylated protein 6 (MPP6) in synapses of mouse cerebrum

doi: 10.1007/s00418-025-02378-1

Figure Lengend Snippet: Western blot analysis of MPP1 ( lanes 1–6 ), MPP2 ( lanes 7–12 ), Lin7 ( lanes 13–18 ), CASK ( lanes 19–24 ), and actin ( lanes 25–30 ) in the cerebrum of Mpp6 +/+ and Mpp6 −/− mice. Samples were obtained from three different Mpp6 +/+ and Mpp6 −/− mice, and their protein concentrations were adjusted as described in the Materials and methods section. Graph showing the relative protein expression ratios of MPP1, MPP2, Lin7, CASK, and actin in Mpp6 −/− mice compared with Mpp6 +/+ mice. Detailed statistical procedures are described in the Materials and Methods section

Article Snippet: The membranes were probed with the following primary antibodies (Abs): rabbit polyclonal anti-MPP6 Abs targeting different regions [N-terminal region (MPP6 Ab no. 1: GeneTex; no. GTX108010), second L27 domain (MPP6 Ab no. 2: ATLAS (Stockholm, Sweden); no. HPA019085), central region (MPP6 Ab no. 3: GeneTex; no. GTX117969), C-terminal region (MPP6 Ab no. 4: originally produced by Saitoh et al. ( )], rabbit polyclonal anti-MPP1 Ab (originally produced by Zhang et al. , rabbit polyclonal anti-MPP2 Ab [GeneTex; no. GTX103908 (Yamada et al. )], rabbit polyclonal anti-Lin7 Ab (GeneTex; no. GTX117114), mouse monoclonal anti-CASK Ab (NeuroMab, Davis, CA, USA; no. K56A/50), mouse monoclonal anti-actin Ab (Wako, Osaka, Japan; clone no. 2F3).

Techniques: Western Blot, Expressing

Journal: Histochemistry and Cell Biology

Article Title: Involvement of membrane palmitoylated protein 6 (MPP6) in synapses of mouse cerebrum

doi: 10.1007/s00418-025-02378-1

Figure Lengend Snippet: Immunolocalization of MPP6 in the mouse cerebrum using light microscopy. a – l : floating sections of the cerebrum, including the cerebral cortex ( Cx : a – c , e – h ), hippocampus ( Hp : a , b , e – h ), amygdala ( Am : i, j ), and corpus putamen ( CPu : k , l ). Approximate coronal Allen Atlas levels 70 ( a – h ), 62 ( i , j ), and 55 ( k , l ) immunostained for MPP6. b , c , d , h , j , and l : higher magnifications of the areas shown in a , b , g , i , and k , respectively. e , f: Control immunostain, without the primary antibody [ f: differential interference contrast (DIC) image]. Am (BL, Ce, and La) basolateral, central, and lateral amygdaloid nucleus, CA cornu Ammonis, CC corpus callosum, Fi fimbria hippocampi, pCx piriform cortex. m – o : confocal laser-scanning microscopic images of MPP6 (red in m , o ) and GluN1 (green in n , o ) in Cx . o is merged image of m and n . The white arrow indicates a red-stained area without green staining, whereas the black arrows indicate areas stained with both red and green. Scale bars : a , e , and f 500 μm; g , i , and k 200 μm; b , c , d , h , j , and l 100 μm; and m – o 2 μm

Article Snippet: The membranes were probed with the following primary antibodies (Abs): rabbit polyclonal anti-MPP6 Abs targeting different regions [N-terminal region (MPP6 Ab no. 1: GeneTex; no. GTX108010), second L27 domain (MPP6 Ab no. 2: ATLAS (Stockholm, Sweden); no. HPA019085), central region (MPP6 Ab no. 3: GeneTex; no. GTX117969), C-terminal region (MPP6 Ab no. 4: originally produced by Saitoh et al. ( )], rabbit polyclonal anti-MPP1 Ab (originally produced by Zhang et al. , rabbit polyclonal anti-MPP2 Ab [GeneTex; no. GTX103908 (Yamada et al. )], rabbit polyclonal anti-Lin7 Ab (GeneTex; no. GTX117114), mouse monoclonal anti-CASK Ab (NeuroMab, Davis, CA, USA; no. K56A/50), mouse monoclonal anti-actin Ab (Wako, Osaka, Japan; clone no. 2F3).

Techniques: Light Microscopy, Control, Staining

Journal: Histochemistry and Cell Biology

Article Title: Involvement of membrane palmitoylated protein 6 (MPP6) in synapses of mouse cerebrum

doi: 10.1007/s00418-025-02378-1

Figure Lengend Snippet: Ultrastructural immunolocalization of MPP6 in the mouse cerebral cortex using post-embedding immunoelectron microscopy. ( a , b ) Light microscopic images of thick sections stained with toluidine blue. ( b ) Higher magnification of a region in a . ( c – i ) Transmission electron micrographs showing 18 nm gold particles conjugated to secondary antibodies localized on and near the postsynaptic densities ( arrows ) following treatment with anti-MPP6 antibody. ( h , i ) Control immunostain without primary anti-MPP6 antibody. Pr presynaptic site, Po postsynaptic site. Scale bars : a 100 μm; b 50 μm; and c – i 50 nm

Article Snippet: The membranes were probed with the following primary antibodies (Abs): rabbit polyclonal anti-MPP6 Abs targeting different regions [N-terminal region (MPP6 Ab no. 1: GeneTex; no. GTX108010), second L27 domain (MPP6 Ab no. 2: ATLAS (Stockholm, Sweden); no. HPA019085), central region (MPP6 Ab no. 3: GeneTex; no. GTX117969), C-terminal region (MPP6 Ab no. 4: originally produced by Saitoh et al. ( )], rabbit polyclonal anti-MPP1 Ab (originally produced by Zhang et al. , rabbit polyclonal anti-MPP2 Ab [GeneTex; no. GTX103908 (Yamada et al. )], rabbit polyclonal anti-Lin7 Ab (GeneTex; no. GTX117114), mouse monoclonal anti-CASK Ab (NeuroMab, Davis, CA, USA; no. K56A/50), mouse monoclonal anti-actin Ab (Wako, Osaka, Japan; clone no. 2F3).

Techniques: Immuno-Electron Microscopy, Staining, Transmission Assay, Control

Journal: Histochemistry and Cell Biology

Article Title: Involvement of membrane palmitoylated protein 6 (MPP6) in synapses of mouse cerebrum

doi: 10.1007/s00418-025-02378-1

Figure Lengend Snippet: Ultrastructural analysis of synapses in the cerebral cortex of Mpp6 +/+ and Mpp6 −/− mice using transmission electron microscopy. ( a , b ) Representative electron micrographs of synapses in Mpp6 +/+ ( a ) and Mpp6 −/− ( b ) mouse cerebral cortex. Arrows indicate the postsynaptic density (PSD). Pr presynaptic site, Po postsynaptic site. ( c ) Schematic representation of a synapse illustrating the synaptic cleft distance (red double-headed arrow) and PSD thickness (blue double-headed arrow) . ( d ) Graph depicting the synaptic cleft distance between presynaptic and postsynaptic membranes ( n = 280 for both Mpp6 +/+ and Mpp6 −/− mice). ( e ) Graph showing PSD thickness ( n = 375 for both Mpp6 +/+ and Mpp6 −/− mice). * p < 0.05, ** p < 0.01. Scale bars: a , b 200 nm

Article Snippet: The membranes were probed with the following primary antibodies (Abs): rabbit polyclonal anti-MPP6 Abs targeting different regions [N-terminal region (MPP6 Ab no. 1: GeneTex; no. GTX108010), second L27 domain (MPP6 Ab no. 2: ATLAS (Stockholm, Sweden); no. HPA019085), central region (MPP6 Ab no. 3: GeneTex; no. GTX117969), C-terminal region (MPP6 Ab no. 4: originally produced by Saitoh et al. ( )], rabbit polyclonal anti-MPP1 Ab (originally produced by Zhang et al. , rabbit polyclonal anti-MPP2 Ab [GeneTex; no. GTX103908 (Yamada et al. )], rabbit polyclonal anti-Lin7 Ab (GeneTex; no. GTX117114), mouse monoclonal anti-CASK Ab (NeuroMab, Davis, CA, USA; no. K56A/50), mouse monoclonal anti-actin Ab (Wako, Osaka, Japan; clone no. 2F3).

Techniques: Transmission Assay, Electron Microscopy

Journal: Histochemistry and Cell Biology

Article Title: Involvement of membrane palmitoylated protein 6 (MPP6) in synapses of mouse cerebrum

doi: 10.1007/s00418-025-02378-1

Figure Lengend Snippet: Elevated plus-maze test. ( a ) Schematic diagram of the apparatus used for the elevated plus-maze test. Detailed construction methods are described in the Materials and methods section. The test began with the mouse placed in the center area. The total time (10 min) spent in the center ( blue area), closed arm (inside the truss bridge; red area), and open arm ( yellow area) was measured for each mouse. ( b ) Graph showing the relative percentage of time spent in different areas (closed arm: red ; center: blue ; and open arm: yellow ) for Mpp6 +/+ and Mpp6 −/− mice at different developmental stages: 2–3 months (2–3 mo.) , 4–7 months (4–7 mo.) , and 18 months (18 mo.)

Article Snippet: The membranes were probed with the following primary antibodies (Abs): rabbit polyclonal anti-MPP6 Abs targeting different regions [N-terminal region (MPP6 Ab no. 1: GeneTex; no. GTX108010), second L27 domain (MPP6 Ab no. 2: ATLAS (Stockholm, Sweden); no. HPA019085), central region (MPP6 Ab no. 3: GeneTex; no. GTX117969), C-terminal region (MPP6 Ab no. 4: originally produced by Saitoh et al. ( )], rabbit polyclonal anti-MPP1 Ab (originally produced by Zhang et al. , rabbit polyclonal anti-MPP2 Ab [GeneTex; no. GTX103908 (Yamada et al. )], rabbit polyclonal anti-Lin7 Ab (GeneTex; no. GTX117114), mouse monoclonal anti-CASK Ab (NeuroMab, Davis, CA, USA; no. K56A/50), mouse monoclonal anti-actin Ab (Wako, Osaka, Japan; clone no. 2F3).

Techniques:

Journal: Histochemistry and Cell Biology

Article Title: Involvement of membrane palmitoylated protein 6 (MPP6) in synapses of mouse cerebrum

doi: 10.1007/s00418-025-02378-1

Figure Lengend Snippet: Locomotor activity test. Detailed methods for measuring activity are described in the Materials and methods section. The graph shows the average locomotor activity counts per hour for Mpp6 +/+ and Mpp6 −/− mice (three mice per group). * p < 0.05, ** p < 0.01, compared at the same times across each hour

Article Snippet: The membranes were probed with the following primary antibodies (Abs): rabbit polyclonal anti-MPP6 Abs targeting different regions [N-terminal region (MPP6 Ab no. 1: GeneTex; no. GTX108010), second L27 domain (MPP6 Ab no. 2: ATLAS (Stockholm, Sweden); no. HPA019085), central region (MPP6 Ab no. 3: GeneTex; no. GTX117969), C-terminal region (MPP6 Ab no. 4: originally produced by Saitoh et al. ( )], rabbit polyclonal anti-MPP1 Ab (originally produced by Zhang et al. , rabbit polyclonal anti-MPP2 Ab [GeneTex; no. GTX103908 (Yamada et al. )], rabbit polyclonal anti-Lin7 Ab (GeneTex; no. GTX117114), mouse monoclonal anti-CASK Ab (NeuroMab, Davis, CA, USA; no. K56A/50), mouse monoclonal anti-actin Ab (Wako, Osaka, Japan; clone no. 2F3).

Techniques: Activity Assay