Journal: American Journal of Physiology - Renal Physiology

Article Title: NBCe1-A is required for the renal ammonia and K + response to hypokalemia

doi: 10.1152/ajprenal.00481.2019

Figure Lengend Snippet: Antibodies used (alphabetical order)

Article Snippet: ENaCβ , StressMarq Biosciences (SPC-404, Victoria, BC, Canada) , WB, 1:5,000.

Techniques: Generated

Journal: bioRxiv

Article Title: Epileptiform activity influences theta-burst induced LTP in the adult hippocampus: a role for lipid raft dynamics in early metaplasticity

doi: 10.1101/2022.12.06.519267

Figure Lengend Snippet: Each panel shows at the bottom the western-blot immunodetection of flotilin-1 ( A. ), caveolin-1 ( B. ), synaptophysin-1 ( C. ) PSD-95 ( D. ), gephyrin ( E. ), and alpha-tubulin ( F. ) obtained in one individual experiment where hippocampal slices underwent Schaffer collateral basal stimulation for 20 min and then were subjected either to interictal-like epileptiform activity (EA) induced by perfusion with 0Mg 2+ for 30 min or ictal-like EA by exposure to bicuculine (10 μM, Bic ) for 16 min, and then allowed to recover for 30 min before slice collection. Control slices were monitored for 70 min (the equivalent time of the EA protocol) before WB analysis. Western blot experiments were performed using synaptosome preparations obtained from these slices. Respective average change in total flotilin-1 ( A. ), caveolin-1 ( B. ), synaptophysin-1 ( C. ) PSD-95 ( D. ), gephyrin ( E. ), and alpha-tubulin ( F. ) immunoreactivities are also plotted at the top in each panel. Individual values and the mean ± S.E.M of five independent experiments are depicted. 100% - averaged PSD-95, gephyrin, caveolin-1, flotillin-1, or synaptophysin-1 immunoreactivity in control conditions (CTR, absence of EA). * P < 0.05 (ANOVA, Sidak’s multiple comparison test) as compared to CTR; ns represents non -significant differences P > 0.05 ( ANOVA , Sidak’s multiple comparison test) between respective bars.

Article Snippet: Membranes were incubated overnight at 4oC with mouse anti-gephyrin (#147011, Synaptic Systems, AB_2810214), rabbit anti-PSD-95 (#CST-2507, Cell Signalling Tech., AB_561221), mouse anti-caveolin-1 (#ab106642, Abcam, AB_10861399), mouse anti-flotillin-1 (#ab133497, Abcam, AB _11156367), rabbit antiphospho-Ser845-GluA1 (1:2500, Abcam #Ab76321; RRID: AB_1523688), rabbit antiphospho-Ser-831-GluA1 (1:2000, Abcam #Ab109464; RRID: AB_10862154), rabbit anti-GluA1 (1:4000, Millipore # AB1504; RRID:AB_2113602), rabbit anti-GluA2 (1:1000, Proteintech #11994-1-AP; RRID: AB_2113725) and rabbit anti-alpha-tubulin (1:5000, Proteintech #11224-1-AP; RRID: AB_2210206) or rabbit anti-beta-actin (1:10000, Proteintech, Cat# 60008-1; RRID:AB_2289225) primary antibodies.

Techniques: Western Blot, Immunodetection, Activity Assay, Control, Comparison

Journal: ACS chemical neuroscience

Article Title: Chronic Social Isolation Stress during Peri-Adolescence Alters Presynaptic Dopamine Terminal Dynamics via Augmentation in Accumbal Dopamine Availability

doi: 10.1021/acschemneuro.8b00360

Figure Lengend Snippet: Assessment of several terminal protein expression levels. Relative protein expression levels of (A) VMAT2 (n = 7 in both groups), (B) Synaptigyrin-3 (aGH: n = 6; aSI: n = 5), (C) Syntaxin-1 (aGH: n = 7; aSI: n = 6), and (D) Munc13-3 (aGH: n = 8; aSI: n = 8) were measured. None of these proteins had different expression levels in aGH and aSI rats. (insets) The representative Western blot images, with the respective protein and actin for comparison. Group housed, aGH, blue; Socially isolated, aSI, red.

Article Snippet: Subsequently, blots were incubated with agitation for 2 h at room temperature in TBS-T/5% bovine serum albumin (05470; Sigma-Aldrich) solution containing the following primary antibody concentrations: VAMT2 (1:2000; AB1598P; Millipore Sigma); Synaptogyrin-3 (1:1000; ab106460; abcam); Syntaxin-1 (1:1000; ANR-002; Alomone laboratories).

Techniques: Expressing, Western Blot, Isolation

Journal: Research and Practice in Thrombosis and Haemostasis

Article Title: SARS-CoV-2 ORF7a activates the endothelium to release von Willebrand factor that promotes thrombosis

doi: 10.1016/j.rpth.2025.102947

Figure Lengend Snippet: Expression and localization of the SARS-CoV-2 ORF7a protein in human embryonic kidney (HEK) 293 cells and splenic vascular endothelial cells (SVECs). (A and B) Immunofluorescent localization of transiently expressed SARS-CoV-2 Orf7a-mCh-flag protein (red) in HEK293 cells and SVECs, respectively. Syntaxin 6 (green) was used for the localization of the Golgi apparatus. 4',6-Diamidino-2-phenylindole stained the nucleus . Scale bar: 10 μm. (C and D) Western blot analysis for detection of the SARS-CoV-2 ORF7a protein in cell lysate and conditioned medium from HEK293 cells and SVECs, respectively. β-actin and α-tubulin were used as the sample loading controls (Ctr.) for HEK293 and SVEC cell lysates, respectively.

Article Snippet: Primary antibodies used were β-actin antibody (Biotechne), recombinant anti–SARS-CoV-2 Orf7a antibody (Abcam), anti-intercellular adhesion molecule 1(Proteintech), anti–α-tubulin monoclonal antibody (Proteintech), and rabbit anti-VWF (Agilent Diagnostics).

Techniques: Expressing, Staining, Western Blot

Journal: Research and Practice in Thrombosis and Haemostasis

Article Title: SARS-CoV-2 ORF7a activates the endothelium to release von Willebrand factor that promotes thrombosis

doi: 10.1016/j.rpth.2025.102947

Figure Lengend Snippet: Flow cytometric detection of the surface expression of von Willebrand factor (VWF) in control (Ctrl) and ORF7a-overexpressing splenic vascular endothelial cells (SVECs). Cell surface VWF was detected by flow cytometry following fixation and staining with Alexa Fluor 647-conjugated anti-VWF immunoglobulin G in the Ctrl cells and SVECs with transient expression of ORF7a. (A) The no-staining negative Ctrl. (B and C) The percentage of cells positive for surface VWF in Ctrl and ORF7a-expressing SVECs, respectively. (D) Quantification of VWF-expressing SVECs without (Ctrl) or with ORF7a transient transfection ( n = 4). Statistical significance was determined by the Student’s t -test, ∗∗ P < .01. FSC-A, forward scatter.

Article Snippet: Primary antibodies used were β-actin antibody (Biotechne), recombinant anti–SARS-CoV-2 Orf7a antibody (Abcam), anti-intercellular adhesion molecule 1(Proteintech), anti–α-tubulin monoclonal antibody (Proteintech), and rabbit anti-VWF (Agilent Diagnostics).

Techniques: Expressing, Control, Flow Cytometry, Staining, Transfection

Journal: Research and Practice in Thrombosis and Haemostasis

Article Title: SARS-CoV-2 ORF7a activates the endothelium to release von Willebrand factor that promotes thrombosis

doi: 10.1016/j.rpth.2025.102947

Figure Lengend Snippet: Soluble SARS-CoV-2 ORF7a protein activates and stimulates von Willebrand factor (VWF) secretion of human endothelial cells. (A) Immunofluorescent detection of intracellular and extracellular distribution of VWF (green) in human umbilical vein endothelial cells (HUVECs) treated with various concentrations (0-2.0 μg/mL) of SARS-CoV-2 ORF7a soluble protein expressed from human embryonic kidney 293 cells for 24 hours. The red arrows indicate elongated VWF fibers or strings, and the pink arrows indicate intracellular VWF staining in Weibel–Palade bodies. 4',6-Diamidino-2-phenylindole (DAPI) stained the nucleus . Scale bar: 10 μm. (B and C) Western blot analyses determined the expression of intercellular adhesion molecule 1 (ICAM1) in primary HUVECs following incubation with different concentrations (0-4.0 μg/mL) of soluble recombinant SARS-CoV-2 ORF7a protein for 24 hours and with a fixed concentration (4.0 μg/mL) of soluble recombinant SARS-CoV-2 ORF7a protein for 0-60 hours, respectively. The α-tubulin protein was used as a sample loading control. (D) Western blot analysis of VWF secreted in the conditioned medium of HUVECs following incubation for 24 hours with a different concentration of the ORF7a protein (0-4.0 μg/mL). A lysine-rich histone (100 μg/mL) was used as a positive stimulator control in all cases.

Article Snippet: Primary antibodies used were β-actin antibody (Biotechne), recombinant anti–SARS-CoV-2 Orf7a antibody (Abcam), anti-intercellular adhesion molecule 1(Proteintech), anti–α-tubulin monoclonal antibody (Proteintech), and rabbit anti-VWF (Agilent Diagnostics).

Techniques: Staining, Western Blot, Expressing, Incubation, Recombinant, Concentration Assay, Control