Journal: iScience

Article Title: The decreased astrocyte-microglia interaction reflects the early characteristics of Alzheimer’s disease

doi: 10.1016/j.isci.2024.109281

Figure Lengend Snippet:

Article Snippet: Primary antibodies used were: anti-Iba1 (1:1000, rabbit, 019–19741, Wako), anti-GFAP (1:1000, rabbit, goat, ab53554, Abcam), anti-6E10 (1:1000, mouse, 803014, biolegends), anti- Synaptophysin (1:1000, rabbit, ab14692, Abcam), anti-Gria1 (1:20000, mouse, 67642-1-IG, proteintech), anti-Gria2 (1:500, rabbit, 11994-1-IG, proteintech), anti-Gria3 (1:1000, rabbit, 29588-1-IG, proteintech), anti-Gria4 (1:500, rabbit, 23350-1-IG, proteintech), anti-PSD95 (1:2000, rabbit, 20665-1-IG, proteintech), anti-Vglut1 (1:1000, rabbit, 12331, Cell signaling), anti-Vim (1:1000, rabbit, YT4880, immunoway), anti-C1qa (1:1000, rabbit, YN0612, immunoway), anti-Lcp1 (1:1000, rabbit, YN0002, immunoway), anti-Anxa3 (1:1000, rabbit, YT0237, immunoway), anti-Anxa5 (1:1000, rabbit, ab14196, Abcam), anti- Ctsb (1:1000, goat, AF965, RD), anti-β-actin (1:5000, mouse, A5441, sigma), Confocal fluorescence images were acquired using a Nikon A1 confocal laser scanning microscope with ×20 objectives for imaging stained or autofluorescent neurons.

Techniques: Virus, Bicinchoninic Acid Protein Assay, Mass Spectrometry, Software

Journal: Frontiers in Molecular Neuroscience

Article Title: Somatic Accumulation of GluA1-AMPA Receptors Leads to Selective Cognitive Impairments in Mice

doi: 10.3389/fnmol.2018.00199

Figure Lengend Snippet: Expression of the L-α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor (AMPAR) subunits and subunit assemblies in WT and Gria1 R/R mice. (A) Hippocampal expression of GluA1–3, GluN1, αCaMKII and ß-actin in WT and Gria1 R/R mice from P2 till P90. (B) Co-immunoprecipitations (IPs) using polyclonal anti-GluA1 and anti-GluA2/3 antibodies show the presence of GluA1–3 in AMPAR assemblies from hippocampal membrane preparations at P > 60 of WT , Gria1 R/R (R/R) and Gria1 −/− (−/−) mice. (C) Schematic representation of the Gria1 R “knock-in” ( Gria1 tm1Erk ) allele and the Gria1 + allele ( WT ). Below the gene segments, the putative AMPAR subtypes, that can operate at CA3-to-CA1 synapses in Gria1 R/R and WT mice, are schematically depicted (GluA1(R) = GluA1(Q600R)). Large AMPAR symbols for high abundance; small symbols for low abundance; transparent for AMPARs with low single channel conductance. The inset shows the position of the Q600R mutations (R) in two out of the four P-loop segments that form the ion pore of an AMPAR. Exons are in boxes, loxP sites in black triangles and the M1 and P-loop coding sequence in black squares. The position of the mutated codon Q600R codon and codon Q600 in Gria1 tm1Erk and Gria1 are indicated, respectively (see Sprengel et al., ). High resolution images of (A,B) are accessible at https://dx.doi.org/10.17617/3.1i .

Article Snippet: The blotted proteins were probed with polyclonal antibodies against GluA1 (Merck Millipore 0.1 μg/ml), anti-GluA2 (Merck Millipore 0.16 μg/ml), anti-GluN1 (Merck Millipore, 0.25 μg/ml, monoclonal anti-GluA3 (ThermoFisher, 1:250), anti-CaMKII (MAB 8699 Merck Millipore 0.1 μg/ml) and anti-ß-actin (AC-15 Ascites, Sigma, 1:25,000), followed by peroxidase-linked anti-rabbit or -mouse secondary antibodies (Jackson Immuno Res., 1:20,000).

Techniques: Expressing, Knock-In, Sequencing