Journal:

Article Title: Muscarinic Acetylcholine Receptor Subtype Expression in Avian Vestibular Hair Cells, Nerve Terminals and Ganglion Cells

doi: 10.1016/j.neuroscience.2007.02.019

Figure Lengend Snippet: Antibodies and Antigens Used for Muscarinic Receptor Subtype Studies

Article Snippet: The primary antibodies included rabbit anti-M1-mAChR polyclonal antibody (Biodesign International, Cat. # Q4A173R, Saco, ME, USA), rabbit anti-M2-mAChR antibody (Alomone Labs Ltd., Cat. # AMR-002, Jerusalem, Israel), rabbit anti-M3-mAChR antibody (Alomone Labs Ltd., Cat. # AMR-006, Jerusalem, Israel), mouse anti-M4-mAChR monoclonal antibody (Chemicon International, Cat. # MAB1578, Temecular, CA) and rabbit anti-M5-mAChR antibody (Biodesign International, Cat. # Q4A871R, Saco, ME, USA).

Techniques:

Journal:

Article Title: Muscarinic Acetylcholine Receptor Subtype Expression in Avian Vestibular Hair Cells, Nerve Terminals and Ganglion Cells

doi: 10.1016/j.neuroscience.2007.02.019

Figure Lengend Snippet: Fluorescent labeling of mAChR subtype expression on vestibular peripheral structures in the crista, utricle and ganglion. The results shown here are the anti-M4 antibody reacted with a red fluorescent secondary antibody while other mAChR subtypes reacted with a green secondary antibody. No differences were found when the red/green fluorescent secondary antibodies were exchanged. The co-expression of M4 with M1, M3 and M5 on nerve fibers, nerve terminals and ganglion cells are marked by arrows. Myelin sheaths and Schwann cells surrounding ganglion cells are pointed at by single arrowheads. Cuticular plates w/ or w/o protruding cilia are pointed at by double arrows. The labeling above the row of hair cells in utricle (especially in E2) is the artifact induced by labeling the otoconial membrane which was not removed. Scale bars = 10 μm.

Article Snippet: The primary antibodies included rabbit anti-M1-mAChR polyclonal antibody (Biodesign International, Cat. # Q4A173R, Saco, ME, USA), rabbit anti-M2-mAChR antibody (Alomone Labs Ltd., Cat. # AMR-002, Jerusalem, Israel), rabbit anti-M3-mAChR antibody (Alomone Labs Ltd., Cat. # AMR-006, Jerusalem, Israel), mouse anti-M4-mAChR monoclonal antibody (Chemicon International, Cat. # MAB1578, Temecular, CA) and rabbit anti-M5-mAChR antibody (Biodesign International, Cat. # Q4A871R, Saco, ME, USA).

Techniques: Labeling, Expressing

Journal:

Article Title: Muscarinic Acetylcholine Receptor Subtype Expression in Avian Vestibular Hair Cells, Nerve Terminals and Ganglion Cells

doi: 10.1016/j.neuroscience.2007.02.019

Figure Lengend Snippet: DAB staining of (A) semicircular canal crista and (B) vestibular ganglion. The number in each panel corresponds to the mAChR subtype number. Myelin sheaths and Schwann cells circumscribe the nerve fibers and ganglion cells (black arrows). Nerve fibers expressing M1, M2, M4 and M5 traverse the basement membrane and terminate in the neuroepithelium (arrowheads). There is no M3 expression on peripheral nerve terminals inside of the neuroepithelium. The myelin sheath staining stops just beneath the basement membrane (double arrowheads in A-3). Scale bars = 10 μm.

Article Snippet: The primary antibodies included rabbit anti-M1-mAChR polyclonal antibody (Biodesign International, Cat. # Q4A173R, Saco, ME, USA), rabbit anti-M2-mAChR antibody (Alomone Labs Ltd., Cat. # AMR-002, Jerusalem, Israel), rabbit anti-M3-mAChR antibody (Alomone Labs Ltd., Cat. # AMR-006, Jerusalem, Israel), mouse anti-M4-mAChR monoclonal antibody (Chemicon International, Cat. # MAB1578, Temecular, CA) and rabbit anti-M5-mAChR antibody (Biodesign International, Cat. # Q4A871R, Saco, ME, USA).

Techniques: Staining, Expressing

Journal:

Article Title: Muscarinic Acetylcholine Receptor Subtype Expression in Avian Vestibular Hair Cells, Nerve Terminals and Ganglion Cells

doi: 10.1016/j.neuroscience.2007.02.019

Figure Lengend Snippet: Summary of mAChR subtype expression on the peripheral vestibular neurons

Article Snippet: The primary antibodies included rabbit anti-M1-mAChR polyclonal antibody (Biodesign International, Cat. # Q4A173R, Saco, ME, USA), rabbit anti-M2-mAChR antibody (Alomone Labs Ltd., Cat. # AMR-002, Jerusalem, Israel), rabbit anti-M3-mAChR antibody (Alomone Labs Ltd., Cat. # AMR-006, Jerusalem, Israel), mouse anti-M4-mAChR monoclonal antibody (Chemicon International, Cat. # MAB1578, Temecular, CA) and rabbit anti-M5-mAChR antibody (Biodesign International, Cat. # Q4A871R, Saco, ME, USA).

Techniques: Expressing

Journal: Scientific reports

Article Title: Transcriptional control of intestinal cholesterol absorption, adipose energy expenditure and lipid handling by Sortilin.

doi: 10.1038/s41598-018-27416-y

Figure Lengend Snippet: Figure 1. Sort1 deficiency reduced body and WAT weight in female Ldlr−/− mice. (a) Body weight in grams (g), (b) body weight gain, and (c) WAT weight ratio for Ldlr−/−Sort1+/+ and Ldlr−/−Sort1−/− mice after 15 weeks (15w) on normal chow (NC) or high-fat/cholesterol diet (HF/HC); (n = 5–10 mice/group). (d) Confirmation of Sortilin protein deficiency in 15-week HF/HC-diet fed female Sort1−/− mice WAT and brown adipose tissue (BAT); (n = 5 mice/group). GAPDH loading controls were run on the same blot, and the membrane was cut prior to incubating with primary antibody. Full-length blots are presented in Supplemental Fig. S4a and b. *P < 0.05, **P < 0.01, ***P < 0.001 versus sex and diet matched Ldlr−/−Sort1+/+ mice, analyzed by t-test; values are presented as mean ± SEM.

Article Snippet: Human cell line Sortilin was detected using an anti-Sortilin antibody (R&D Systems #AF3154) and GAPDH (Santa Cruz, #sc-47724, Dallas, TX, USA) was used as a loading control.

Techniques: Membrane

Journal: Scientific reports

Article Title: Transcriptional control of intestinal cholesterol absorption, adipose energy expenditure and lipid handling by Sortilin.

doi: 10.1038/s41598-018-27416-y

Figure Lengend Snippet: Figure 6. SORT1 deficiency suppressed LXR-mediated transcription in human Caco-2, HepG2, and Hek293 cells. (a) Sortilin protein reduction confirmation in Caco-2, HepG2, and Hek293 cells following SORT1 RNA interference (siSort1) or scrambled control (Scr), with or without 24-hour 1 μM T0901317 (T090) LXR agonist treatment; (n = 3; representative western blots shown). GAPDH loading controls were run on the same blot, and the membrane was cut prior to incubating with primary antibody. Full-length blots are presented in Supplemental Fig. S4c. (b) Caco-2, (c) HepG2, and (d) Hek293 cells SORT1, LXRα/β (NR1H3, NR1H2), and KLF4 mRNA levels following SORT1 RNA interference, with or without 24-hour 1 μM T090 treatment; (n = 3–6). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 vs. Scr control, analyzed by ANOVA with Tukey’s multiple comparisons test; values are presented as mean ± SEM.

Article Snippet: Human cell line Sortilin was detected using an anti-Sortilin antibody (R&D Systems #AF3154) and GAPDH (Santa Cruz, #sc-47724, Dallas, TX, USA) was used as a loading control.

Techniques: Control, Western Blot, Membrane

Journal: Function

Article Title: Calcium Signaling in Pancreatic Immune Cells In situ

doi: 10.1093/function/zqaa026

Figure Lengend Snippet: Immunostaining after Recording ATP-elicited Ca 2+ Signals in pancreatic macrophages (PMs). (A). Representative images of pancreatic lobule loaded with Fluo-4AM before (Ai) and after ATP (10 µM) application (Aii), the arrow indicates the position of the ( n = 8). A corresponding Fluo 4 trace from a PM is shown in Aiii . Corresponding immunostaining of this lobule with antibodies F4/80 Alexa Fluo 647 is shown below ( Aiv) . Hoechst 33342 staining of the same area is shown in Av . Arrow points to ear-like shape of PM nucleus. Overlay of antibody and Hoechst 33342 staining is shown in Avi . Scale bar is 10µm. (B). Immunostaining of another area in a pancreatic lobule with monoclonal F4/80 antibodies labeled with Alexa Fluor 647 (Bi) . Staining of nuclei in the same lobule with Hoechst 33342 (Bii) . Overlay of B i with Bii is shown in Biii . Scale bar is 10µm. (C). Representative images of a pancreatic lobule loaded with Fluo-4AM before (Ci) and after ATP (10 µM) application (Cii) , the arrow indicates the position of the PM. Corresponding Fluo 4 trace is shown in Ciii. Immunostaining of the same area with monoclonal CD11b antibody conjugated with Alexa Fluor 647 ( n = 8) is shown in Civ . Overlay of Cii and Civ is shown in Cv . Scale bar is 10µm.

Article Snippet: Mouse F4/80 monoclonal rat Antibody (CI-A3-1) [Alexa Fluor ® 647] and mouse CD11b/Integrin alpha M Alexa Fluor ® 647-conjugated monoclonal rat antibodies were obtained from Novus Biologicals Europe and R&D Systems Bio-techne, respectively.

Techniques: Immunostaining, Staining, Labeling

Journal: Function

Article Title: Calcium Signaling in Pancreatic Immune Cells In situ

doi: 10.1093/function/zqaa026

Figure Lengend Snippet: IgG-elicited Ca 2+ Spikes in PMs . (A). Single short Ca 2+ spike occurring after application of IgG (0.1–0.25 mg/mL) in a PM from a control pancreatic lobule. This was an infrequent observation (5 out of 29 cells tested) and is most likely not an IgG-elicited Ca 2+ signal as such single spikes have been also observed in 3 out of 15 cells in the absence of IgG stimulation. (B) . Representative trace of IgG (0.1–0.25 mg/mL)-induced Ca 2+ signals in PMs in pancreatic lobules isolated from mice with AP (FAEE-AP model—48 h). Such oscillations were observed in 9 out of 31 cells. Single short spikes have been observed in 4 out of 31 cells. No oscillations were observed in the absence of stimulation with IgG ( n = 14), while single short spikes have been observed in 2 out of 14 cells. (C). Average Ca 2+ spike frequencies in PMs displaying Ca 2+ signals under the conditions indicated. The frequencies in control PMs, both stimulated with IgG (blue bar) and unstimulated (green), as well as in unstimulated PMs from the FAEE-AP model (48 h, orange bar) were much lower than in PMs from the FAEE-AP model stimulated with IgG (red bar, P < 0.007). (D) . Average Ca 2+ spike duration in PMs displaying Ca 2+ signals under the conditions indicated. Although the average spike duration was longer in the PMs from the FAEE-AP mice stimulated with IgG than under the other conditions, the difference was not statistically different ( P > 0.2). (E). Representative images of immunostaining of PMs in lobules using antibodies F4/80 conjugated with Alexa Fluor 647. Lobules were isolated from control and FAEE-AP 3-day mice (72 h in vivo FAEE-AP model). Scale bar is 20µm. (F). Comparison of the average density of PMs in lobules from control and FAEE-AP 2-day and 3-day mice (48 h and 72 h in vivo FAEE-AP model, respectively). Control, 2.36 ± 0.6 SEM, n = 14; FAEE-AP 2 day, 9.56 ± 1.86 SEM, * P < 0.033, n = 16; FAEE-AP 3 days, 15.37 ± 1.51 SEM, * P < 0.038 as compared to FAEE-AP 2-day, n = 35. The difference between control and FAEE-AP 3-day was very highly significant (**** P < 0.0001).

Article Snippet: Mouse F4/80 monoclonal rat Antibody (CI-A3-1) [Alexa Fluor ® 647] and mouse CD11b/Integrin alpha M Alexa Fluor ® 647-conjugated monoclonal rat antibodies were obtained from Novus Biologicals Europe and R&D Systems Bio-techne, respectively.

Techniques: Isolation, Immunostaining, In Vivo, Comparison