Journal: Physiological Reports

Article Title: Tuberous sclerosis complex exhibits a new renal cystogenic mechanism

doi: 10.14814/phy2.13983

Figure Lengend Snippet: List of antibodies used in the study

Article Snippet: Rabbit polyclonal Anti‐NCC , 1:60 , Stressmarq Biosciences; Victoria, BC, Canada , SPC‐402.

Techniques: Plasmid Preparation, Generated

Journal: Frontiers in Molecular Neuroscience

Article Title: Behavioral and Molecular Responses to Exogenous Cannabinoids During Pentylenetetrazol-Induced Convulsions in Male and Female Rats

doi: 10.3389/fnmol.2022.868583

Figure Lengend Snippet: Expression levels of proteins obtained from a fraction enriched in cytoplasmic proteins/proteins bound to small organelles. (A) Representative Western blots of CB1R, GluA2R, βArr2, PSD95, p-CaMKII and total CaMKII in females and (I) males. (B–H) Quantification of (A) (Females. CB1R: C, n = 5; PTZ, n = 8; WIN, n = 4; WIN + PTZ, n = 6; GluA2: C, n = 5; PTZ, n = 9; WIN, n = 5; WIN + PTZ, n = 6; βArr2: C, n = 5; PTZ, n = 9; WIN, n = 5; WIN + PTZ, n = 6; PSD95: C, n = 5; PTZ, n = 8; WIN, n = 5; WIN + PTZ, n = 6, p -CaMKIIα: C, n = 5; PTZ, n = 8; WIN, n = 4; WIN + PTZ, n = 5; p -CaMKIIβ: C, n = 5; PTZ, n = 7; WIN, n = 5; WIN + PTZ, n = 5; total CaMKII: C, n = 3; PTZ, n = 4; WIN, n = 3; WIN + PTZ, n = 3). (J–P) Quantification of (D) (Males. CB1R: C, n = 4; PTZ, n = 6; WIN, n = 3; WIN + PTZ, n = 5; GluA2: C, n = 4; PTZ, n = 7; WIN, n = 5; WIN + PTZ, n = 5; βArr2: C, n = 4; PTZ, n = 5; WIN, n = 4; WIN + PTZ, n = 5; PSD95: C, n = 4; PTZ, n = 6; WIN, n = 4; WIN + PTZ, n = 5, p -CaMKIIα: C, n = 4; PTZ, n = 5; WIN, n = 4; WIN + PTZ, n = 5; p -CaMKIIβ: C, n = 4; PTZ, n = 5; WIN, n = 4; WIN + PTZ, n = 5; total CaMKII: C, n = 3; PTZ, n = 4; WIN, n = 3; WIN + PTZ, n = 3). Tubulin was used as protein loading control for all gels. * p < 0.05 and ** p < 0.01, one-way ANOVA with Tukey’s multiple comparisons post hoc .

Article Snippet: Primary antibodies used were anti CB1, 1:1,000 (Ab23703, Abcam, UK), anti βArr2, 1:500 (Ab31294, Abcam), anti phospho T286phosphorylated calcium/calmodulin-dependent protein kinase II (p-CaMKII), 1:2,000 (Ab32678, Abcam), anti postsynaptic density protein 95 (PSD95) 1:1,000 (#610496, BD Biosciences, USA), anti α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid, AMPA, subunit GluA2 (GluA2), 1:1,000 (Sc-517265, Santa Cruz Biotechnology, USA), anti calcium/calmodulin-dependent protein kinase II (total CaMKII) (Sc-5306, Santa Cruz Biotechnology), antiGolgi matrix protein 130 (GM130), 1:1,000 (#610823, BD Biosciences, USA), anti βIII-Tubulin, 1:2,500 (G7121, Promega, USA), anti N-cadherin, 1:1,000 (#33-3900, Invitrogen).

Techniques: Expressing, Western Blot, Control

Journal: Investigative Ophthalmology & Visual Science

Article Title: Essential Role of XBP1 in Maintaining Photoreceptor Synaptic Integrity in Early Diabetic Retinopathy

doi: 10.1167/iovs.64.14.40

Figure Lengend Snippet: Confocal microscopy of ribbon synapses labeled with Ribeye and mGluR6 in mouse retinas. Cryosections (12 µm thick) of mouse retinas were stained with antibodies against metabotropic glutamate receptor 6 (mGluR6, red ) and Ribeye ( green ) to label the presynaptic (Ribeye) and postsynaptic (mGluR6) components of ribbon synapses between photoreceptors and bipolar cells in the OPL. Ribeye also extensively labeled synapses within the IPL. Nuclei were stained with DAPI ( blue ). ( A – D ) Representative confocal images from five mice per group. Each image is a projection of confocal scans taken from five consecutive focal planes. Scale bar : 50 µm.

Article Snippet: After blocking, sections were incubated with primary antibodies including anti-GABA (PC213L, 1:800, Millipore Sigma, St. Louis, Missouri, USA), anti-Ribeye (anti-ctbp2, 612044, 1:600 for QUANTOS, BD Biosciences, Franklin Lakes, New Jersey, USA; anti-Ribeye, 192 003, 1:600 for PSD-95 colabeling, Synaptic Systems, Göettingen, Germany), anti-mGluR6 (AGC-026, 1:600, Alomone, Jerusalem, Israel), and anti-PSD95 (MA1046, 1:600, ThermoFisher, Waltham, Massachusetts, USA) antibodies, followed by secondary antibodies (A11001, T6391, A11015, at 1:800, Invitrogen, Carlsbad, CA, USA).

Techniques: Confocal Microscopy, Labeling, Staining

Journal: Journal of Neuropathology and Experimental Neurology

Article Title: Friedreich Ataxia: Developmental Failure of the Dorsal Root Entry Zone

doi: 10.1093/jnen/nlx087

Figure Lengend Snippet: DREZ in a normal control and a patient with FA shown by α-dystroglycan immunohistochemistry and double-label immunofluorescence of periaxin and GFAP. (A, B) Normal control; FA (C, D) (FA2, Table, GAA 1200/1200). (A) In the normal control, compact α-dystroglycan reaction product surrounds the dome of the CNS portion of the DREZ. (B) Double-label immunofluorescence of periaxin and GFAP shows a relatively sharp transition of myelinating Schwann cells (red) to GFAP-reactive CNS tissue (green) in the DREZ. (C) In FA, the demarcation between α-dystroglycan-positive and negative regions of the DREZ is interrupted by short breaks where CNS tissue penetrates into the DR (arrows). (D) In FA, the normal demarcation between Schwann cells (periaxin) and CNS (GFAP) is indistinct. (A, C) α-dystroglycan; (B, D) double-label immunofluorescence of periaxin (red) and GFAP (green). Scale bars: (A, C) 200 µm; (B, D) 100 µm.

Article Snippet: Sources of commercial antibodies; catalog numbers; host species and clonality; final protein concentrations or, in case of ascites fluid, dilution; and antigen retrieval methods were as follows: anti-PMP-22 (Abcam, Cambridge, MA, ab-31851, rabbit polyclonal, 2 μg protein/mL, incubation in 80% ethanol at 4 °C overnight); anti-P0 (Abcam, ab15508, rabbit polyclonal, 1 μg protein/mL, incubation in 80% ethanol at 4 °C overnight, followed by 0.01 M citric acid-sodium citrate buffer, pH 6, for 20 minutes at 95 °C); anti-PLP (Abcam, ab-105784, rabbit polyclonal, 2 μg protein/mL, incubation in 80% ethanol at 4 °C overnight, followed by 0.01 M citric acid-sodium citrate buffer, pH 6, for 20 minutes at 95 °C); anti-α-dystroglycan (Novus Biologicals, Littleton, CO, NBP1-49634, mouse monoclonal, 0.4 μg protein/mL, 0.01 M citric acid-sodium citrate buffer, pH 6, for 20 minutes at 95 °C); anti-periaxin (Novus, NBP1-89598, rabbit polyclonal, 2 μg protein/mL, 0.01 M citric acid-sodium citrate buffer, pH 6, 20 minutes at 95 °C); anti-laminin (Sigma-Aldrich, St. Louis, MO, L8271, mouse monoclonal, ascites fluid, diluted 1:1000, proteinase K [Sigma, P-6556], 0.1 μg/mL in tris buffer, pH 8, containing 0.01 M NaCl for 30 minutes at 37 °C); anti-GFAP (BioLegend, San Diego, CA, 837201, mouse monoclonal, ascites fluid, diluted 1:500, DIVA [a proprietary decloaking solution sold by Biocare Medical, Pacheco, CA] 1× for 30 minutes at 95 °C); anti-vimentin (Santa Cruz Biotechnology, Santa Cruz, CA, sc-66002, mouse monoclonal, 2 μg protein/mL, 0.01 M citric acid-sodium citrate buffer, pH 6, 20 minutes at 95 °C); anti-nestin (Santa Cruz, sc-23927, mouse monoclonal, 2 μg protein/mL, 0.01 M citric acid-sodium citrate buffer, pH 6, for 20 minutes at 95 °C, or DIVA for 30 minutes at 95 °C, or trypsin [DIFCO Laboratories, Detroit, MI, 0152-13] 1 mg/mL in 0.07 M CaCl 2 at pH 7.8 for 30 minutes at 37 °C); anti-S100 (Santa Cruz, sc-53438, mouse monoclonal, 0.8 μg protein/mL, 0.01 M citric acid-sodium citrate buffer, pH 6, 20 minutes at 95 °C); anti-class-III-β-tubulin (R&D Systems, Minneapolis, MN, MAB1195, mouse monoclonal, 2 μg protein/mL, 0.01 M citric acid-sodium citrate buffer, pH 6, 20 minutes at 95 °C); anti-phosphorylated neurofilament protein (Covance [now BioLegend] SMI-31 R, mouse monoclonal, ascites fluid, diluted 1:500, 0.01 M citric acid-sodium citrate buffer, pH 6, 20 minutes at 95 °C).

Techniques: Immunohistochemistry, Immunofluorescence