Journal: bioRxiv

Article Title: Dynamic translocation of Inside-Out proteins to the cell surface underlies cellular adaptation to cancer-induced stress

doi: 10.1101/2025.10.30.685532

Figure Lengend Snippet: (A) PC3 and OVCAR3 cells were subjected to real-time monitoring of anti-Myl6 IgG internalization using the Incucyte S3 live-cell analysis system. Quantitative analysis of red fluorescence over time reveals a continuous, time-dependent increase in anti-Myl6 IgG uptake, while the isotype control antibody exhibits no detectable internalization (n = 3, mean ± SD). (B) Confocal microscopy images of HCT116 live-cell immunostaining performed at 4°C or 37°C. An absence of anti-Myl6 IgG internalization at 4°C is demonstrated by fluorescence signal restricted to the cell surface, whereas prominent intracellular accumulation is observed at 37°C, confirming active, temperature-dependent uptake. Scale bar denotes 10 μ m.

Article Snippet: PC3 cells were seeded at a density of 10,000 cells per well in black-walled 96-well plates (Thermo Scientific) and incubated overnight to allow for adherence. α-Myl6 IgG and isotype control IgG were employed at a final concentration of 4 μg/mL and conjugated with the Incucyte Human Fab-fluor-pH Antibody Labeling Dye for Antibody Internalization (Red) (Sartorius) according to the manufacturer’s protocol.

Techniques: Cell Analysis, Fluorescence, Control, Confocal Microscopy, Immunostaining

Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

Article Title: Prematurity reduces citrulline-arginine-nitric oxide production and precedes the onset of necrotizing enterocolitis in piglets

doi: 10.1152/ajpgi.00198.2018

Figure Lengend Snippet: Multiple end-point measurements of necrotizing enterocolitis (NEC) incidence and severity in piglets. A: Kaplan-Meir graph detailing piglet survival. B: gross NEC score in piglets of advancing gestational age exposed to the NEC protocol. A threshold of 3 in any one segment was considered positive for NEC (dashed line). C: histological NEC score was determined in hematoxylin-eosin-stained tissue sections, and a threshold of ≥2 in any tissue section was considered positive for NEC (dashed line). D: plasma intestinal fatty-acid binding protein (iFABP) concentration in the final plasma sample for piglet groups in response to the NEC protocol. B–D: *P < 0.05, significant differences between groups, and error bars show 95% confidence interval. Data analyzed by Kruskal-Wallis test and Dunn’s multiple comparison test. NS, not significant.

Article Snippet: Amino acid concentrations in plasma and feed were measured using phenyl isothiocyanate derivatives relative to a methionine sulfone internal standard by HPLC ( 6 ). iFABP was measured using a human iFABP Quantikine ELISA Kit (R&D Systems).

Techniques: Staining, Clinical Proteomics, Binding Assay, Concentration Assay, Comparison

Journal: Journal of Histochemistry and Cytochemistry

Article Title: A Flexible Mouse-On-Mouse Immunohistochemical Staining Technique Adaptable to Biotin-Free Reagents, Immunofluorescence, and Multiple Antibody Staining

doi: 10.1369/0022155413511620

Figure Lengend Snippet: Protocol Steps in Sequential Order Depending on the Secondary Antibody.

Article Snippet: To create an antibody complex, we combined a biotinylated goat anti-mouse IgG Fab fragment (#115-067-003; Jackson ImmunoResearch Laboratories; West Grove, PA), the desired mouse primary antibody and antibody diluent in a tube at room temperature for 20 min.

Techniques: Blocking Assay, Polymer

Journal: Journal of Histochemistry and Cytochemistry

Article Title: A Flexible Mouse-On-Mouse Immunohistochemical Staining Technique Adaptable to Biotin-Free Reagents, Immunofluorescence, and Multiple Antibody Staining

doi: 10.1369/0022155413511620

Figure Lengend Snippet: Mouse-on-mouse staining method on mouse kidney (A, B, C), spleen (D, E, F), and intestine (G, H, I) tissue samples. Panels (A), (D), and (G) illustrate the amount of staining from endogenous mouse immunoglobulin if only anti-mouse secondary antibody is added to tissue. Panels (C), (F), and (I) are stained with the anti- smooth muscle actin (SMA) antibody complexed with the biotinylated secondary antibody. Panels (B), (E), and (H) are stained with an isotype control antibody complexed with the secondary antibody at the same concentration as the anti-SMA antibody. Images were captured at 10×. Scale bar is 100 µm.

Article Snippet: To create an antibody complex, we combined a biotinylated goat anti-mouse IgG Fab fragment (#115-067-003; Jackson ImmunoResearch Laboratories; West Grove, PA), the desired mouse primary antibody and antibody diluent in a tube at room temperature for 20 min.

Techniques: Staining, Control, Concentration Assay