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

Journal: iScience

Article Title: Spatiotemporally organized immunomodulatory response to SARS-CoV-2 virus in primary human broncho-alveolar epithelia

doi: 10.1016/j.isci.2023.107374

Figure Lengend Snippet:

Article Snippet: Anti-cytokeratin 5 AF594 (Polyclonal) , Novus , CatNBP2-61931AF594 RRID: AB_2933967.

Techniques: Bacteria, Virus, Variant Assay, Recombinant, Red Blood Cell Lysis, Software, Membrane, Cell Culture

Journal: Nanomaterials

Article Title: Inhibition of TRPV1 Channel Activity in Human CD4 + T Cells by Nanodiamond and Nanoplatinum Liquid, DPV576

doi: 10.3390/nano8100770

Figure Lengend Snippet: ( A – D ) DPV576 does not modulate the expression of TRPV4 on CD4 + T cells. Inactivated and anti-CD3/CD28 activated CD4 + T lymphocytes were exposed to DPV576 for 24 h. The cells were stained for TRPV channels. Expression of TRPV4 on non-activated T cells ( A , B ). Anti-CD3/CD28 activated T cells ( C , D ). Data is representative of three experiments. Blue: Without DPV576; Red: DPV576 exposed lymphocytes; Green: Isotype.

Article Snippet: Briefly, cells were collected and centrifuged, re-suspended in PBS (phosphate buffered saline) 2% FBS, and incubated with specific antibodies for TRPV1-AL647 (bs-1931R-A647) or TRPV4-Alexa 488 (bs-6425R-A488) (Bioss Inc., Woburn, MA, USA) at a dilution of 1:100 for 1 h. Subsequently, the cells were washed and a minimum of 10,000 cells were acquired on FACS Calibur (Becton Dickinson, San Jose, CA, USA).

Techniques: Expressing, Staining

Journal: Cell

Article Title: HIF Regulates Multiple Translated Endogenous Retroviruses: Implications for Cancer Immunotherapy

doi: 10.1016/j.cell.2025.01.046

Figure Lengend Snippet: Key resource table

Article Snippet: Mouse monoclonal IgG2 a AF647 , Santa Cruz Biotechnology , Cat#sc-24637, RRID: AB_737235.

Techniques: Virus, Recombinant, SYBR Green Assay, cDNA Synthesis, Purification, Sample Prep, Sequencing, Amplification, Software, Binding Assay, Transferring, Reverse Transcription, Low Protein Binding, Antibody Purification, Adjuvant, Enzyme-linked Immunospot