Journal: Neuron

Article Title: APP Family Regulates Neuronal Excitability and Synaptic Plasticity but Not Neuronal Survival

doi: 10.1016/j.neuron.2020.08.011

Figure Lengend Snippet: (A) Schematic illustrations of APP, APLP1, and APLP2 showing conserved protein structure and domains. APP695 is the abundant isoform derived from exons 1–6, 9–18; APLP1654 is derived from exons 1–17; APLP2751 is derived from exons 1–13, 15–18 (based on Ensembl Genome Database: APP, ENSMUSG00000022892; APLP1, ENSMUSG00000006651; APLP2, ENSMUSG00000031996); SP: signaling peptide; E1: the extracellular domain containing the heparin-binding motif, the growth factor-like domain, and the cooper- and zinc-binding motif; Ac: an acidic domain; KPI: the Kunitz-type protease inhibitor domain; E2: the extracellular domain containing the heparin-binding motif and the random coiled region; Aβ: the amyloid beta peptide; Y: the YENPTY motif. (B) Schematic illustrations of the gene structures of APP, APLP1, and APLP2. The blue (APP), green (APLP1), and orange (APLP2) boxes represent exons encoding the protein sequences. The gray boxes represent the 5’ and 3’ UTRs. The exons are not drawn to scale, and exon 1 contains the start codon ATG. The promoter and the first exon (for APP and APLP2) or the first two exons (for APLP1) are flanked by two loxP sites (red arrowheads). (C) Reduced levels of the APP, APLP1, or APLP2 mRNA in the cortex of cKO mice at 2–3 months of ages, relative to controls. The remaining mRNA detected in the cortex of cKO mice is likely due to APP, APLP1, or APLP2 expression in interneurons and glia, where the Camk2a-Cre transgene is not expressed. (D) Dramatic decreases of APP, APLP1 and APLP2 proteins in the neocortex (NCX) of cTKO mice at the age of 3 months. (E-J”) Immunohistochemical analysis shows abundant APP immunoreactivity (green), which co-localizes with NeuN immunoreactivity (red), in the NCX and hippocampal area CA1 (CA1) of control mice, whereas APP immunoreactivity is largely eliminated in the NCX (F) and hippocampal area CA1 (H) of cTKO mice. Most GAD67-immunoreactive (red) interneurons in the NCX of control mice appear to co-express APP (I”), and the few APP-immunoreactive cells in the NCX of cTKO mice appear to be co-labeled with GAD67 immunoreactivity (J”). Scale bar: 100 μm.

Article Snippet: Primary antibodies used were rabbit anti-APP (Sigma-Aldrich, A8717, RRID: AB_258409), rabbit anti-APP-Y188 (Abcam, ab32136, RRID: AB_2289606), rabbit anti-APLP1 (Millipore, #171615, RRID: AB_565288), rabbit anti-APLP1 (CT11, gift of D. Walsh), rabbit anti-APLP2 (Millipore, #171616, RRID: AB_211446), rabbit anti-APLP2 (D2-II, gift of D. Walsh), rabbit anti-MAP2 (Cell Signaling, #4542, RRID: AB_776174), rabbit anti-SYP (Cell Signaling, #5431, RRID: AB_10698743), rabbit anti-SNAP25 (abcam, ab108990, RRID: AB_10888111), rabbit anti-PSD95 (Cell Signaling, #2507, RRID: AB_1264242), anti-GFAP (Sigma-Aldrich, G6171, RRID: AB_1840893), mouse anti-β-actin (Sigma-Aldrich, A1978, RRID: AB_476692), mouse anti-β-actin (Cell Signaling, #3700, RRID: AB_2242334), and rabbit anti-β-actin (Cell Signaling, #4967, RRID: AB_330288).

Techniques: Derivative Assay, Binding Assay, Protease Inhibitor, Expressing, Immunohistochemical staining, Labeling

Journal: Neuron

Article Title: APP Family Regulates Neuronal Excitability and Synaptic Plasticity but Not Neuronal Survival

doi: 10.1016/j.neuron.2020.08.011

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Primary antibodies used were rabbit anti-APP (Sigma-Aldrich, A8717, RRID: AB_258409), rabbit anti-APP-Y188 (Abcam, ab32136, RRID: AB_2289606), rabbit anti-APLP1 (Millipore, #171615, RRID: AB_565288), rabbit anti-APLP1 (CT11, gift of D. Walsh), rabbit anti-APLP2 (Millipore, #171616, RRID: AB_211446), rabbit anti-APLP2 (D2-II, gift of D. Walsh), rabbit anti-MAP2 (Cell Signaling, #4542, RRID: AB_776174), rabbit anti-SYP (Cell Signaling, #5431, RRID: AB_10698743), rabbit anti-SNAP25 (abcam, ab108990, RRID: AB_10888111), rabbit anti-PSD95 (Cell Signaling, #2507, RRID: AB_1264242), anti-GFAP (Sigma-Aldrich, G6171, RRID: AB_1840893), mouse anti-β-actin (Sigma-Aldrich, A1978, RRID: AB_476692), mouse anti-β-actin (Cell Signaling, #3700, RRID: AB_2242334), and rabbit anti-β-actin (Cell Signaling, #4967, RRID: AB_330288).

Techniques: Plasmid Preparation, Recombinant, Protease Inhibitor, Blocking Assay, Electron Microscopy, In Situ, Transgenic Assay, Software

Journal: Oncology Letters

Article Title: Role of APLP2 in the prognosis and clinicopathology of renal cell carcinoma

doi: 10.3892/ol.2018.9577

Figure Lengend Snippet: mRNA expression of APLP2 in normal and CCRCC tissues, as detected by reverse transcription-quantitative polymerase chain reaction. (A) APLP2 mRNA levels in 8 pairs of tissues. (B) Significantly decreased expression of APLP2 in CCRCC tissues (P=0.02). CCRCC, clear cell renal cell carcinoma; APLP2, amyloid β precursor-like protein 2. *P<0.05, **P<0.01, compared with normal.

Article Snippet: The PVDF membrane was blocked for 1 h at room temperature with 5% non-fat milk with Tris-buffered saline Tween-20 (TBST) dilution and then incubated with the aforementioned rabbit anti-APLP2 antibody (1:1,000 dilution) or β-actin antibody (1:1,000 dilution; cat. no. BM0627; Wuhan Boster Biological Technology, Ltd.) overnight at 4°C.

Techniques: Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Oncology Letters

Article Title: Role of APLP2 in the prognosis and clinicopathology of renal cell carcinoma

doi: 10.3892/ol.2018.9577

Figure Lengend Snippet: APLP2 protein expression in clear cell renal cell carcinoma and matched normal kidney samples. T, tumor sample; N, matched normal sample; APLP2, amyloid β precursor-like protein 2.

Article Snippet: The PVDF membrane was blocked for 1 h at room temperature with 5% non-fat milk with Tris-buffered saline Tween-20 (TBST) dilution and then incubated with the aforementioned rabbit anti-APLP2 antibody (1:1,000 dilution) or β-actin antibody (1:1,000 dilution; cat. no. BM0627; Wuhan Boster Biological Technology, Ltd.) overnight at 4°C.

Techniques: Expressing

Journal: Oncology Letters

Article Title: Role of APLP2 in the prognosis and clinicopathology of renal cell carcinoma

doi: 10.3892/ol.2018.9577

Figure Lengend Snippet: Expression of APLP2 protein observed by immunohistochemical staining. The arrows point to cells with a positive staining of APLP2. (A) Expression of APLP2 protein in normal kidney tissues (score >6). (B) Expression of APLP2 protein in CCRCC tissues (score <6). (C) Expression of APLP2 protein in CCRCC tissues (score >6). (D) Expression of APLP2 protein in normal kidney tissues (score <6). Magnification, ×200. CCRCC, clear cell renal cell carcinoma; APLP2, amyloid β precursor-like protein 2.

Article Snippet: The PVDF membrane was blocked for 1 h at room temperature with 5% non-fat milk with Tris-buffered saline Tween-20 (TBST) dilution and then incubated with the aforementioned rabbit anti-APLP2 antibody (1:1,000 dilution) or β-actin antibody (1:1,000 dilution; cat. no. BM0627; Wuhan Boster Biological Technology, Ltd.) overnight at 4°C.

Techniques: Expressing, Immunohistochemical staining, Staining

Journal: Oncology Letters

Article Title: Role of APLP2 in the prognosis and clinicopathology of renal cell carcinoma

doi: 10.3892/ol.2018.9577

Figure Lengend Snippet: Clinicopathological features and their associations with APLP2 expression.

Article Snippet: The PVDF membrane was blocked for 1 h at room temperature with 5% non-fat milk with Tris-buffered saline Tween-20 (TBST) dilution and then incubated with the aforementioned rabbit anti-APLP2 antibody (1:1,000 dilution) or β-actin antibody (1:1,000 dilution; cat. no. BM0627; Wuhan Boster Biological Technology, Ltd.) overnight at 4°C.

Techniques: Expressing

Journal: Oncology Letters

Article Title: Role of APLP2 in the prognosis and clinicopathology of renal cell carcinoma

doi: 10.3892/ol.2018.9577

Figure Lengend Snippet: Kaplan-Meier survival curves of APLP2 expression in clear cell renal cell carcinoma tissues. Patients with low expression of APLP2 exhibited reduced disease-specific survival rates, compared with patients with high expression of APLP2. P<0.001. APLP2, amyloid β precursor-like protein 2.

Article Snippet: The PVDF membrane was blocked for 1 h at room temperature with 5% non-fat milk with Tris-buffered saline Tween-20 (TBST) dilution and then incubated with the aforementioned rabbit anti-APLP2 antibody (1:1,000 dilution) or β-actin antibody (1:1,000 dilution; cat. no. BM0627; Wuhan Boster Biological Technology, Ltd.) overnight at 4°C.

Techniques: Expressing

Journal: Oncology Letters

Article Title: Role of APLP2 in the prognosis and clinicopathology of renal cell carcinoma

doi: 10.3892/ol.2018.9577

Figure Lengend Snippet: Cox regression analyses for disease-specific survival.

Article Snippet: The PVDF membrane was blocked for 1 h at room temperature with 5% non-fat milk with Tris-buffered saline Tween-20 (TBST) dilution and then incubated with the aforementioned rabbit anti-APLP2 antibody (1:1,000 dilution) or β-actin antibody (1:1,000 dilution; cat. no. BM0627; Wuhan Boster Biological Technology, Ltd.) overnight at 4°C.

Techniques: