Journal: Dentistry Journal

Article Title: Investigation of the Molecular Profile of Granular Cell Tumours and Schwannomas of the Oral Cavity

doi: 10.3390/dj10030038

Figure Lengend Snippet: Summary of the antibodies used for immunohistochemistry of GCTs and schwannomas.

Article Snippet: GAP43 , Bio-Techne Canada, Oakville On, NB300-143 , Rabbit polyclonal , Regenerating neural tissues/growth cones, GAP43 intracellular growth protein/membrane protein , 1/5000.

Techniques: Immunohistochemistry, Cell Surface Receptor Assay, Membrane, Plasmid Preparation

Journal: Diabetes

Article Title: Loss of NADPH Oxidase–Derived Superoxide Skews Macrophage Phenotypes to Delay Type 1 Diabetes

doi: 10.2337/db14-0929

Figure Lengend Snippet: NOD.Ncf1m1J islets exhibit lowered M1 and elevated M2 levels of macrophage markers. Purified islet whole-cell lysates from prediabetic 16-week-old NOD and NOD.Ncf1m1J mice were used in an immunoblot analysis for P-STAT6 (Y641) and STAT6 (A), iNOS (B), and Arg-1 (C). Densitometry statistics for Western blotting represent the average of three experiments. ***P < 0.001; *P < 0.05. WT, wild type.

Article Snippet: For Western blotting, whole-cell lysates were probed with antibodies against iNOS (Novus Biologicals), Arg-1, STAT6 (Cell Signaling), P-STAT6 (Y641) (BD Bioscience), or β-actin (Sigma-Aldrich) as described ( 18 ).

Techniques: Purification, Western Blot

Journal: Scientific Reports

Article Title: Cervical spinal cord stimulation exerts anti-epileptic effects in a rat model of epileptic seizure through the suppression of CCL2-mediated cascades

doi: 10.1038/s41598-024-64972-y

Figure Lengend Snippet: ( A , C ) Double immunofluorescence staining of DAPI (blue) with Iba-1 (green) and GFAP (red) in the hippocampal CA1, CA3, and DG 7 days after the administration of KA. Scale bar = 80 µm. ( B , D ) Quantitative data of d showing the number of Iba1- and GFAP-positive cells in the hippocampus. Data are presented as the mean ± standard error. n = 7 rats for each group. * p < 0.05, ** p < 0.01, *** p < 0.001. Abbreviations: Iba-1, ionized calcium-binding adapter molecule 1; GFAP, glial fibrillary acidic protein; DAPI, 4,6-diamidino-2-phenylindole; DG, dentate gyrus; KA, kainic acid.

Article Snippet: The following primary antibodies were used for tissue staining: rabbit anti-Iba1 antibody (1:250; Wako Pure Chemical Industries, Osaka, Japan), rabbit anti-GFAP antibody (1:1000; Novus Biologicals, Littleton, CO).

Techniques: Double Immunofluorescence Staining, Binding Assay

Journal: JCI Insight

Article Title: A metabolic redox relay supports ER proinsulin export in pancreatic islet β cells

doi: 10.1172/jci.insight.178725

Figure Lengend Snippet: Male and female 12- to 16-week-old mouse islets ( A – C ) or INS-1 832/3 cells ( D and E ) were treated with vehicle (veh) or mannoheptulose (MnH; 2 mM or 1 mM, respectively) for 4 hours as indicated. NADPH/NADP + ( A , n = 4) and GSSG/GSH ( B , n = 4) were measured by sequential incubation in 2 mM Glc followed by 20 mM Glc for 12 minutes each via ratiometric imaging of iNAP or Grx1-roGFP2 (AdRIP), respectively. Responses were normalized to 2 mM Glc. ( C ) ER redox was measured ( n = 5) via ratiometric imaging of ERroGFP (AdRIP). ( D and E ) INS-1 832/3 cells stably expressing proCpepSNAP cells were treated with veh or MnH or cotreated with MnH plus DTT (0.5 mM; MnH + DTT) for 4 hours as indicated. Cells were pulse-labeled with SNAP-505 (green), chased for 10 minutes, immunostained for TGN38 (magenta) and BiP (red), and counterstained with DAPI (blue). ( D ) The ratio of proCpepSNAP fluorescence coincident with the Golgi (TGN38) versus ER (BiP) was quantified ( n = 4). ( E ) Representative images are shown. ( A – D ) Data represent the mean ± SEM. * P < 0.05 by 2-tailed Student’s t test ( A – C ) or 1-way ANOVA with Tukey’s posttest analysis ( D ). Scale bar = 5 μm.

Article Snippet: For immunostaining, cells were incubated overnight with antibodies raised against BiP (rabbit; gift of Christopher Nicchitta, Duke University, Durham, North Carolina, USA; 1:200), GRASP55 (rabbit, Proteintech 10598-AP, 1:500), GM130 (mouse, BD Transduction 610-823, 1:200), TGN38 (mouse, Novus Biologicals, Bio-Techne, NB300-575, 1:200), or proinsulin (mouse, MyBioSource MBS660187, 1:200) as indicated.

Techniques: Incubation, Imaging, Stable Transfection, Expressing, Labeling, Fluorescence

Journal: JCI Insight

Article Title: A metabolic redox relay supports ER proinsulin export in pancreatic islet β cells

doi: 10.1172/jci.insight.178725

Figure Lengend Snippet: ( A and B ) Male and female 12- to 16-week-old mouse islets or INS-1 832/3 cells were treated with vehicle (veh), 2-AAPA (25 μM), or auranofin (AFN; 10 μM) for 4 hours before imaging. ( A ) NADPH/NADP + were measured in islets ( n = 4) by sequential incubation in 2 mM Glc followed by 20 mM Glc for 12 minutes each via ratiometric imaging of iNAP (AdRIP). Responses were normalized to 2 mM Glc. ( B ) INS-1 832/3 cells stably expressing proCpepSNAP were pulse-labeled with SNAP-505, chased for 10 minutes, immunostained for BiP and TGN38, and counterstained with DAPI. The ratio of proCpepSNAP fluorescence coincident with the Golgi (TGN38) versus the ER (BiP) was quantified ( n = 3). ( C – G ) Male and female 12- to 16-week-old mouse islets were treated with Ad-shSAFE or Ad-sh Txnrd1 as indicated and analyzed 96 hours after infection. ( C ) Txnrd1 mRNA expression was quantified by RT-qPCR ( n = 3–4). ( D , E , and G ) Male and female 12- to 16-week-old mouse islets expressing proCpepSNAP (AdRIP) were pulse-labeled with SNAP-505 (green) and chased for 10 minutes. Cells were fixed, immunostained for GM130 (magenta), and counterstained for DAPI (blue). Representative images are shown ( D ) and the ratio of proCpepSNAP coincident with the Golgi (GM130) versus non-Golgi region in mCherry + cells ( n = 5) was quantified ( E ) and total fluorescence intensity calculated ( G ). ( F ) ER redox was measured in mCherry + islet cells (Ad-shRNA; n = 4) via ratiometric imaging of ERroGFP (AdRIP). ( A – C and E – G ) Data represent the mean ± SEM. * P < 0.05 by 1-way ANOVA with Dunnett’s posttest analysis ( A ), 1-way ANOVA with Tukey’s posttest analysis ( B ), or 2-tailed Student’s t test ( C and E – G ). Scale bar = 5 μm.

Article Snippet: For immunostaining, cells were incubated overnight with antibodies raised against BiP (rabbit; gift of Christopher Nicchitta, Duke University, Durham, North Carolina, USA; 1:200), GRASP55 (rabbit, Proteintech 10598-AP, 1:500), GM130 (mouse, BD Transduction 610-823, 1:200), TGN38 (mouse, Novus Biologicals, Bio-Techne, NB300-575, 1:200), or proinsulin (mouse, MyBioSource MBS660187, 1:200) as indicated.

Techniques: Imaging, Incubation, Stable Transfection, Expressing, Labeling, Fluorescence, Infection, Quantitative RT-PCR, shRNA