Journal: iScience

Article Title: Differential conservation analysis identifies residues defining constitutive internalization in beta-adrenergic receptors

doi: 10.1016/j.isci.2026.115033

Figure Lengend Snippet: B2AR V34A 1.33 and S41A 1.40 mutations alter the organization of receptor homomers (A) Schematic showing the experimental design of BRET for receptor-receptor associations. HEK 293 cells were transiently transfected with equal amounts of Rluc8-tagged receptor, either wild-type or with mutations, and increasing amounts of Venus-tagged receptor, either wild-type or with mutations. BRET saturation curves were used to determine the (B) BRET max and (C) the BRET 50. Data are the mean ± SEM; one-sample t test: ∗ p < 0.05. (D) Curve plotted using one-site specific binding saturation equation, representative curves shown, n = 6–8.

Article Snippet: HEK 293 , ATCC , CRL-1573; RRID:CVCL_0045.

Techniques: Transfection, Binding Assay

Journal: iScience

Article Title: Differential conservation analysis identifies residues defining constitutive internalization in beta-adrenergic receptors

doi: 10.1016/j.isci.2026.115033

Figure Lengend Snippet: The effect of B2AR mutations on basal and ligand-induced cAMP (A) cAMP accumulation in HEK 293 cells transiently transfected with HA-B2AR WT and mutants with isoproterenol treatment (1 pM-10 μM, 5-min). (B) cAMP production at the maximum isoproterenol dose (10 μM) and (C) basal cAMP production. Data are the mean ± SEM from n = 5–6 independent experiments. One-sample t test: ∗∗∗ p < 0.001. (D) EC 50 values for curves in A.

Article Snippet: HEK 293 , ATCC , CRL-1573; RRID:CVCL_0045.

Techniques: Transfection

Journal: iScience

Article Title: Differential conservation analysis identifies residues defining constitutive internalization in beta-adrenergic receptors

doi: 10.1016/j.isci.2026.115033

Figure Lengend Snippet: Double mutation decreases basal activity by increasing constitutive internalization (A and B) Cell surface expression of HEK 293 cells transiently transfected with HA-B2AR WT or mutant receptors. (A) Percent cells expressing receptor shown as fold change to the WT and (B) the amount of receptor at the plasma membrane shown as fold change to the WT. n = 5, one sample t test: ∗ p < 0.05, ∗∗ p < 0.01. (C) Whole-cell expression of B2AR-Rluc8 mutant receptors shown as fold change to the WT receptor, n = 6–10. One-sample t test: ∗∗ p < 0.01. (D and E) cAMP production in (D) basal and (E) isoproterenol (1 pM-10 μM, 5-min) treated HEK 293 cells transiently transfected with WT and V34A-S41A B2AR and matched for receptor expression at the plasma membrane . (F) Schematic demonstrating antibody conditions for labeling endocytic/constitutively internalized receptor pools and the total pool of receptor (endocytic + biosynthetic). (G) Confocal microscopy images of HEK 293 cells transiently transfected with HA-B2AR WT and mutant receptors, either fed live with anti-HA antibody (endocytic imaging) or incubated with anti-HA antibody post-fixation and permeabilization (endocytic + biosynthetic imaging). Representative images shown, n = 10 cells. Scale bars, 10 μm; inset = 3 μm. (H) Constitutive internalization measured in HEK 293 cells transiently transfected with HA-B2AR, HA-V34A-S41A B2AR, or HA-B1AR live labeled with anti-HA antibody at 4°C with/without 1-h incubation at 37°C. Cell surface expression measured via flow cytometry. Data shown as percentage change from surface expression in cells kept at 4°C, mean ± SEM. n = 4. One-way ANOVA with Šídák’s multiple comparisons test: ∗ p < 0.05; ∗∗ p < 0.01. See also .

Article Snippet: HEK 293 , ATCC , CRL-1573; RRID:CVCL_0045.

Techniques: Mutagenesis, Activity Assay, Expressing, Transfection, Clinical Proteomics, Membrane, Labeling, Confocal Microscopy, Imaging, Incubation, Flow Cytometry

Journal: bioRxiv

Article Title: Oncostatin M orchestrates collective epithelial migration via HIF1A activation

doi: 10.1101/2025.09.26.678830

Figure Lengend Snippet: (A) MCF10A cells were treated with EGF, OSM, or IFNG for 48 hours, then stained for DAPI (blue) and β-Catenin (red). (B) Cell velocity vectors represent speed and direction of cell motility, derived from cell tracking data during 24-48 hours after ligand treatment. Arrow size is proportional to velocity. (C-F) Quantification of cell phenotype from live-cell imaging. Shaded region represents 95% confidence interval from 3 biological replicates (n=3). Boxplots in (E) depict interquartile range of nearest neighbor distance calculated for each cell. (G-J) Line plots show protein expression levels normalized to the T0 control for each treatment condition. Error bars represent the full range of measurements (n=3).

Article Snippet: Afterward, cells were treated with either 10 ng/ml EGF (R&D Systems #236-EG), 10 ng/ml OSM (R&D Systems #8475-OM), or 20 ng/ml IFNG (R&D Systems #258-IF) + 10 ng/mL EGF.

Techniques: Staining, Derivative Assay, Cell Tracking Assay, Live Cell Imaging, Expressing, Control

Journal: bioRxiv

Article Title: Oncostatin M orchestrates collective epithelial migration via HIF1A activation

doi: 10.1101/2025.09.26.678830

Figure Lengend Snippet: (A) Workflow for the comparative network analysis to identify molecular subnetworks and nodes perturbed by OSM. Integrated molecular data collected from OSM and IFNG treated cells was analyzed using CausalPath. (B) Combined molecular network of OSM and IFNG activated nodes. The rewiring score between conditions is indicated by node color, and the top scoring nodes nominated for further investigation are indicated by increased size. (C) The top scoring nodes most rewired by OSM treatment, corresponding to the highlighted nodes in (B). (D) The majority (8/14) of top rewired nodes are centered around the STAT3 subnetwork.

Article Snippet: Afterward, cells were treated with either 10 ng/ml EGF (R&D Systems #236-EG), 10 ng/ml OSM (R&D Systems #8475-OM), or 20 ng/ml IFNG (R&D Systems #258-IF) + 10 ng/mL EGF.

Techniques: