Journal: Frontiers in Synaptic Neuroscience

Article Title: Akap5 links synaptic dysfunction to neuroinflammatory signaling in a mouse model of infantile neuronal ceroid lipofuscinosis

doi: 10.3389/fnsyn.2024.1384625

Figure Lengend Snippet: Key resources table.

Article Snippet: Antibody , Mouse monoclonal anti-GluN2B , UC Davis/NIH NeuroMab Facility , Cat: 75/097; RRID: AB_10673405 , 1:1,000.

Techniques: Recombinant, Software

Journal: Brain Communications

Article Title: Presenilin1 familial Alzheimer disease mutants inactivate EFNB1- and BDNF-dependent neuroprotection against excitotoxicity by affecting neuroprotective complexes of N -methyl- d -aspartate receptor

doi: 10.1093/braincomms/fcaa100

Figure Lengend Snippet: Effects of EFNB1 and PS1 FAD mutants on NMDAR interactions with PS1. ( A ) Cortical neuronal cultures were stimulated with EFNB1 (eB1) or Fc for 60 min. After stimulation, cells were lysed and IPed with anti-PS1 NTF antibody, then immunoblotted with anti-GLUN1 and anti-GLUN2B antibodies. PS1 NTF (PS1) was detected under the same condition with unboiled samples. Graphs show fold change of GLUN1 immunoprecipitated with PS1 following eB1 stimulation (right panel). Experiments were repeated at least three times. Data are mean ± SEM. * P < 0.05 versus Fc, two-tailed paired t -test, n = 3. The inputs are shown in the lower panel. ( B ) Cortical neuronal cultures from WT, heterozygous (KI/WT) and homozygous (KI/KI) for PS1 FAD KI mutant M146V or I213T mouse embryonic brains were treated with eB1 or Fc for 60 min, lysed and IPed with anti-PS1 NTF antibody, then immunoblotted with anti-GLUN1. PS1 was detected under the same condition with unboiled samples. PIS = pre-immune serum. ( C ) Three-month-old WT and PS1 FAD (M146V and I213T) KI heterozygous (KI/WT) or homozygous (KI/KI) mouse brain cortices were homogenized in Hepes buffer containing 1% Triton-X 100. After centrifugation, total lysates were IPed with anti-PS1 NTF antibody, then immunoblotted with anti-GLUN1 and anti-GLUN2B antibodies. PS1 was detected under the same condition with unboiled samples.

Article Snippet: Antibodies used in our studies were as follows: for IP experiments; anti-PS1 (rabbit polyclonal, R222) ( Georgakopoulos et al. , 1999 ), anti-EPHB2 (rabbit polyclonal, R407) ( Barthet et al. , 2013 ), anti-GLUN1 (mouse monoclonal; Novus Biologicals, Littleton, CO, USA) and anti-TRKB (rabbit polyclonal, Abcam; Cambridge, MA, USA); for western blot (WB) detection; anti-PS1 (rabbit polyclonal, R222), anti-PS1 (mouse monoclonal, 33B10) ( Huang et al. , 2018 ), anti-EPHB2 (rabbit polyclonal, Zymed; San Francisco, CA, USA), anti-GLUN1 (mouse monoclonal; BD Biosciences, San Jose, CA, USA), anti-GLUN2B (mouse monoclonal; BD Biosciences, San Jose, CA, USA), anti-ACTIN (mouse monoclonal; Abcam, Cambridge, MA, USA), anti-TRKB (rabbit polyclonal, Abcam; Cambridge, MA, USA); for immunofluorescent experiments; anti-EPHB2 (rabbit polyclonal, Abcam; Cambridge, MA, USA), anti-GLUN1 (mouse monoclonal; Biolegend, San Diego, CA, USA) and anti-PS1 (rabbit polyclonal, R222, affinity purified).

Techniques: Immunoprecipitation, Two Tailed Test, Mutagenesis, Centrifugation