Journal: bioRxiv

Article Title: Arp2/3 complex-dependent actin remodeling is required for efficient RSV uncoating in A549 cells

doi: 10.64898/2026.03.11.711013

Figure Lengend Snippet: (A) Representative confocal microscopy images of RSV-GFP (MOI = 1) bound to A549 WT and Arp2 KO17 cells. Cells were incubated with virus or mock-infected for 1 h on ice, washed to remove unbound virus, fixed, and stained with Palivizumab (anti-RSV-fusion protein, in pseudocolor cyan), Hoechst 33342 (cell nuclei, in pseudocolor green), and WGA (wheat germ agglutinin-Alexa Flour 647, magenta). Images were acquired on a spinning-disk microscope and are shown as maximum-intensity projections of respective channels. Scale bar, 50 µm. (B) Quantification of viral binding from the image series in (A). Pooled data of three individual experiments (three biological replicates per experiment). n denotes the number of cells analysed. Experiments are color-coded; each dot represents one image, and each rectangle represents the experiment mean. Mean ± SD. ns, not significant (Two-way ANOVA). (C) Immunoblot of endogenous IGF1R and nucleolin in A549 WT and Arp2 KO cells. GAPDH (glyceraldehyde-3-phosphate dehydrogenase) served as loading control. (D) Schematic of using PALM in combination with TIRF to assess IGF1R diffusion and clustering. This image was created using Biorender (License number: CT29AQJ52X). (E) Diffusion coefficient of A549 WT and Arp2 KO cells stably expressing IGF1R-mEos3.2. The data were obtained from sptPALM imaging. ns, not significant (unpaired two-tailed Student’s t-test). (F) Representative sptPALM reconstruction with tracks of individual IGF1R-mEos3.2 particles generated from 10,000 frames (frame interval 30 ms). Trajectories are color-coded. (G) Comparison of the IGF1R cluster size of A549 WT and Arp2 KO cells stably expressing IGF1R-mEos3.2. ns, not significant (unpaired two-tailed Student’s t-test). (H) Representative PALM image showing the surface distribution of IGF1R-mEos3.2 in an A549 WT cell.

Article Snippet: The antibodies used in this study were as follows: rabbit recombinant monoclonal anti-Arp2 antibody (1:2,000; EPR7979, ab129018, Abcam); mouse monoclonal anti-HSC70 antibody (1:5,000; sc-7298, Santa Cruz); GAPDH (D16H11) rabbit monoclonal antibody (1:1,000, 5174S, Cell Signaling Technology); IGF1R polyclonal antibody (1:500, BS-0227R, Bioss); nucleolin monoclonal antibody (1:500, 39-6400, Invitrogen); anti-Beta lactamase antibody (1:500, ab12251, Abcam); anti-SARS-CoV-2 nucleocapsid protein antibody (1:500, ABK79-E11-M, Abcalis); Palivizumab (10 μg/mL, a gift from Dr. Thomas Pietschmann, TWINCORE, Germany); HRP-conjugated goat anti-rabbit antibody (1:10,000; G-21234, Invitrogen); HRP-conjugated goat anti-mouse antibody (1:10,000; G-21040, Invitrogen); goat anti-Human cross-adsorbed secondary antibody, Alexa FluorTM 488 (1:200, A-11013, Invitrogen); goat anti-human IgG cross-adsorbed secondary antibody, HRP (1:10,000; A18811, Invitrogen).

Techniques: Confocal Microscopy, Incubation, Virus, Infection, Staining, Microscopy, Binding Assay, Western Blot, Control, Diffusion-based Assay, Stable Transfection, Expressing, Imaging, Two Tailed Test, Generated, Comparison

Journal: iScience

Article Title: The proprotein convertase furin regulates the development of thymic epithelial cells to ensure central immune tolerance

doi: 10.1016/j.isci.2022.105233

Figure Lengend Snippet: Furin promoted the development and proliferation of TECs via proteolytic cleavage of proIGF1R and proIR (A) A row-normalized heatmap showing the expression of gene pathways that gradually decreased in mTECs and cTECs of 2-week-old WT, Furin fl/+ Foxn1 Cre , and Furin fl/fl Foxn1 Cre mice. (B) UMAP plots (left) and violin plots (right) showing the expression of Igf1r and Insr on mTECs and cTECs of 2-week-old WT and Furin fl/fl Foxn1 Cre mice. (C) Immunoblotting of cell lysates of cultured WT and Furin KO TECs for detecting the cleavage of proIGF1R and proIR. (D) Immunoblotting of cell lysates of cultured WT and Furin KO TECs for detecting the phosphorylation of IGF1R/IR, ERK1/2, and Akt (Thr308 and Ser473). (E) mTECs of WT and Furin fl/fl Foxn1 Cre mice were sorted by flow cytometry, and cell lysates were analyzed by Simple Western to detect the cleavage of proIGF1R and proIR. (F) Cultured WT and Furin KO TECs were starved overnight following stimulation with exogenous IGF1 (100 ng/mL) and insulin (200 ng/mL) for 20 min, and cell lysates were analyzed by immunoblot for detecting the phosphorylation of IGF1R/IR, ERK1/2, and Akt (Thr308 and Ser473). In panels C-F, the results are representative of three independent experiments. (G) The numbers of mTECs and cTECs in ERK inhibitor (SCH772984) treated mice (n = 4) and control mice (n = 5). (H) The numbers of mTECs and cTECs in Akt inhibitor (MK-2206) treated fetal thymic lobes (n = 6) and control fetal thymic lobes (n = 7). Data are represented as mean ± SD ∗p < 0.05; ∗∗p < 0.01; NS, not statistically different.

Article Snippet: The primary antibodies used for Simple Western are as follows: β-Actin (Sigma-Aldrich, A5441), IGF1R antibody (Novus Biologicals, NBP1-77679), and Insulin receptor β (CST, 3025).

Techniques: Expressing, Western Blot, Cell Culture, Phospho-proteomics, Flow Cytometry, Simple Western, Control

Journal: iScience

Article Title: The proprotein convertase furin regulates the development of thymic epithelial cells to ensure central immune tolerance

doi: 10.1016/j.isci.2022.105233

Figure Lengend Snippet:

Article Snippet: The primary antibodies used for Simple Western are as follows: β-Actin (Sigma-Aldrich, A5441), IGF1R antibody (Novus Biologicals, NBP1-77679), and Insulin receptor β (CST, 3025).

Techniques: Plasmid Preparation, Recombinant, Reverse Transcription, Bicinchoninic Acid Protein Assay, Software

Journal: Journal of Molecular Endocrinology

Article Title: Human aortic smooth muscle cells are insulin resistant at the receptor level but sensitive to IGF1 and IGF2

doi: 10.1677/jme-09-0021

Figure Lengend Snippet: Figure 1 Expression of IR and IGF1R mRNA and proteins in HASMC. The relative amount of IGF1R to IR mRNA expression was measured by real-time RT-PCR, where IR was set to 1. The IGF1R and IR protein levels were analyzed by ELISA and are expressed as amount receptor protein relative to amount of total protein, IR was set to 1. The IGF1R to IR mRNA ratio (5.3G1.3) and the IGF1R to IR protein ratio (5.5G1.2) was found to be almost identical. ***Denotes P!0.001 (meanGS.E.M., nZ4).

Article Snippet: After washing away the unbound fraction with PBS–T, the secondary antibody was added and incubated on a shaker at RT for 2 h. The biotinylated anti-human IGF1R antibody (BAF 391; R&D Systems) was diluted as 0.5 mg/l in 0.1 w/v % HSA in PBS.

Techniques: Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: Journal of Molecular Endocrinology

Article Title: Human aortic smooth muscle cells are insulin resistant at the receptor level but sensitive to IGF1 and IGF2

doi: 10.1677/jme-09-0021

Figure Lengend Snippet: Figure 3 Detection of IR and IGF1R b-subunits in HASMC. IR and IGF1R b-subunit protein and evidence for the presence of hybrid IR/IGF1R were demonstrated by immunoprecipitation and western blot. Insulin receptor (IR) b-subunit (95 kDa) and IGF1R (IGF1R) b-subunit (97 kDa) were detected by western blot on the same membrane implying co-precipitation. Figures show blots after immunoprecipitation (IP) against either IR (left column, A and B), or IGF1R (right column, C and D) and immunoblot against the IR b-subunit (A and D) and the IGF1R b-subunit (B and C). Similar results were obtained in four independent experiments.

Article Snippet: After washing away the unbound fraction with PBS–T, the secondary antibody was added and incubated on a shaker at RT for 2 h. The biotinylated anti-human IGF1R antibody (BAF 391; R&D Systems) was diluted as 0.5 mg/l in 0.1 w/v % HSA in PBS.

Techniques: Immunoprecipitation, Western Blot, Membrane

Journal: Journal of Molecular Endocrinology

Article Title: Human aortic smooth muscle cells are insulin resistant at the receptor level but sensitive to IGF1 and IGF2

doi: 10.1677/jme-09-0021

Figure Lengend Snippet: Figure 4 (A) Activation of IR b-subunit in HASMC. Effect of insulin (dark gray bars), IGF1 (black bars), and IGF2 (gray bars) on tyrosine phosphorylation of IR b-subunit were analyzed by immunoprecipitation of IR by an anti-IR b-subunit antibody and immunoblotting with an anti-phosphotyrosine antibody (PY20) after exposure of near confluent HASMC to insulin, IGF1, and IGF2 at the concentrations of 10K10–10K8 mol/l for 10 min. Data are means from four independent experiments. Data were quantified by densitometry and are expressed as phosphor- ylated/total protein, fractional of maximal response. *P!0.05. (B) Activation of IGF1R b-subunit in HASMC. Effects of insulin (dark gray bars), IGF1 (black bars), and IGF2 (gray bars) on tyrosine phosphorylation of IGF1R b-subunit were analyzed by immunoprecipitation of IGF1R by an anti IGF1R b-subunit antibody and immunoblotting with an anti-phosphotyrosine antibody (PY20) after exposure of near confluent HASMC to insulin, IGF1, and IGF2 at the concentrations of 10K10– 10K8 mol/l for 10 min. Data are means from four independent experiments. Data were quantified by densitometry and are expressed as phosphorylated/total protein, fractional of maximal response. *P!0.05.

Article Snippet: After washing away the unbound fraction with PBS–T, the secondary antibody was added and incubated on a shaker at RT for 2 h. The biotinylated anti-human IGF1R antibody (BAF 391; R&D Systems) was diluted as 0.5 mg/l in 0.1 w/v % HSA in PBS.

Techniques: Activation Assay, Phospho-proteomics, Immunoprecipitation, Western Blot

Journal: Journal of Molecular Endocrinology

Article Title: Human aortic smooth muscle cells are insulin resistant at the receptor level but sensitive to IGF1 and IGF2

doi: 10.1677/jme-09-0021

Figure Lengend Snippet: Figure 8 Proposed model for insulin resistance in HASMC. The insulin resistance of vascular smooth muscle is located at the receptor level. Due to the low number of IRs there are not enough receptors activated by insulin to generate a downstream signal. However, the IR b-subunit is activated by IGF1 when incorporated into hybrid IR/IGF1R. Both IGF1 and IGF2 activate the IGF1R and the hybrid IR/IGF1R and this elicits downstream signaling and biological effects.

Article Snippet: After washing away the unbound fraction with PBS–T, the secondary antibody was added and incubated on a shaker at RT for 2 h. The biotinylated anti-human IGF1R antibody (BAF 391; R&D Systems) was diluted as 0.5 mg/l in 0.1 w/v % HSA in PBS.

Techniques:

Journal: PLOS ONE

Article Title: Insulin-like growth factor 1 receptor expression correlates with programmed death ligand 1 expression and poor survival in non-small cell lung cancer

doi: 10.1371/journal.pone.0297397

Figure Lengend Snippet: (A) Immunostaining for IGF1R reveals predominant cytoplasmic staining. (B) Immunostaining for PD-L1 reveals predominant cytoplasmic staining. (C) Hematoxylin-eosin staining.

Article Snippet: Following this, the slides were blocked with 10% normal mouse serum for 10 min at room temperature and incubated overnight at 4°C with the anti-IGF1R antibody (NB110-87052, 1:200; Novus Biologicals LLC, Centennial, CO, USA).

Techniques: Immunostaining, Staining

Journal: PLOS ONE

Article Title: Insulin-like growth factor 1 receptor expression correlates with programmed death ligand 1 expression and poor survival in non-small cell lung cancer

doi: 10.1371/journal.pone.0297397

Figure Lengend Snippet: Patient characteristics.

Article Snippet: Following this, the slides were blocked with 10% normal mouse serum for 10 min at room temperature and incubated overnight at 4°C with the anti-IGF1R antibody (NB110-87052, 1:200; Novus Biologicals LLC, Centennial, CO, USA).

Techniques: Adjuvant

Journal: PLOS ONE

Article Title: Insulin-like growth factor 1 receptor expression correlates with programmed death ligand 1 expression and poor survival in non-small cell lung cancer

doi: 10.1371/journal.pone.0297397

Figure Lengend Snippet: Multivariate analysis of the relationship between IGF1R expression and PD-L1 expression.

Article Snippet: Following this, the slides were blocked with 10% normal mouse serum for 10 min at room temperature and incubated overnight at 4°C with the anti-IGF1R antibody (NB110-87052, 1:200; Novus Biologicals LLC, Centennial, CO, USA).

Techniques: Expressing

Journal: PLOS ONE

Article Title: Insulin-like growth factor 1 receptor expression correlates with programmed death ligand 1 expression and poor survival in non-small cell lung cancer

doi: 10.1371/journal.pone.0297397

Figure Lengend Snippet: (A) Comparison of smoking index between the IGF1R-positive and negative groups. The IGF1R-positive group had a significantly larger smoking index. (B) Comparison of smoking index between PD-L1-positive and negative groups. The PD-L1-positive group had a significantly larger smoking index. (C) Comparison of maximum tumor diameter between IGF1R-positive and negative groups. The IGF1R-positive group had a significantly larger maximum tumor diameter.

Article Snippet: Following this, the slides were blocked with 10% normal mouse serum for 10 min at room temperature and incubated overnight at 4°C with the anti-IGF1R antibody (NB110-87052, 1:200; Novus Biologicals LLC, Centennial, CO, USA).

Techniques: Comparison

Journal: PLOS ONE

Article Title: Insulin-like growth factor 1 receptor expression correlates with programmed death ligand 1 expression and poor survival in non-small cell lung cancer

doi: 10.1371/journal.pone.0297397

Figure Lengend Snippet: (A) Kaplan-Meier curve depicting RFS for all patients. The IGF1R-positive group exhibited significantly worse RFS. (B) Kaplan-Meier curve depicting OS for all patients. The IGF1R-positive group exhibited significantly worse OS.

Article Snippet: Following this, the slides were blocked with 10% normal mouse serum for 10 min at room temperature and incubated overnight at 4°C with the anti-IGF1R antibody (NB110-87052, 1:200; Novus Biologicals LLC, Centennial, CO, USA).

Techniques:

Journal: PLOS ONE

Article Title: Insulin-like growth factor 1 receptor expression correlates with programmed death ligand 1 expression and poor survival in non-small cell lung cancer

doi: 10.1371/journal.pone.0297397

Figure Lengend Snippet: (A) Kaplan-Meier curve depicting RFS in stage 0–1 patients. The IGF1R-positive group exhibited significantly worse RFS. (B) Kaplan-Meier curve depicting OS in stage 0–1 patients. The IGF1R-positive group exhibited significantly worse OS.

Article Snippet: Following this, the slides were blocked with 10% normal mouse serum for 10 min at room temperature and incubated overnight at 4°C with the anti-IGF1R antibody (NB110-87052, 1:200; Novus Biologicals LLC, Centennial, CO, USA).

Techniques:

Journal: PLOS ONE

Article Title: Insulin-like growth factor 1 receptor expression correlates with programmed death ligand 1 expression and poor survival in non-small cell lung cancer

doi: 10.1371/journal.pone.0297397

Figure Lengend Snippet: (A) Kaplan-Meier curve depicting RFS in stage 2 patients. No significant difference was observed between the IGF1R-positive and negative groups. (B) Kaplan-Meier curve for OS in stage 2 patients. No significant difference was observed between the IGF1R-positive and negative groups.

Article Snippet: Following this, the slides were blocked with 10% normal mouse serum for 10 min at room temperature and incubated overnight at 4°C with the anti-IGF1R antibody (NB110-87052, 1:200; Novus Biologicals LLC, Centennial, CO, USA).

Techniques:

Journal: PLOS ONE

Article Title: Insulin-like growth factor 1 receptor expression correlates with programmed death ligand 1 expression and poor survival in non-small cell lung cancer

doi: 10.1371/journal.pone.0297397

Figure Lengend Snippet: (A) Kaplan-Meier curve depicting RFS in stage 3 patients. No significant difference in RFS was observed between the IGF1R-positive and negative groups. (B) Kaplan-Meier curve depicting OS in stage 3 patients. No significant difference was observed between the IGF1-positive and negative groups.

Article Snippet: Following this, the slides were blocked with 10% normal mouse serum for 10 min at room temperature and incubated overnight at 4°C with the anti-IGF1R antibody (NB110-87052, 1:200; Novus Biologicals LLC, Centennial, CO, USA).

Techniques:

Journal: PLOS ONE

Article Title: Insulin-like growth factor 1 receptor expression correlates with programmed death ligand 1 expression and poor survival in non-small cell lung cancer

doi: 10.1371/journal.pone.0297397

Figure Lengend Snippet: Multivariable analysis of RFS in stage 0–1 patients.

Article Snippet: Following this, the slides were blocked with 10% normal mouse serum for 10 min at room temperature and incubated overnight at 4°C with the anti-IGF1R antibody (NB110-87052, 1:200; Novus Biologicals LLC, Centennial, CO, USA).

Techniques: Adjuvant

Journal: PLOS ONE

Article Title: Insulin-like growth factor 1 receptor expression correlates with programmed death ligand 1 expression and poor survival in non-small cell lung cancer

doi: 10.1371/journal.pone.0297397

Figure Lengend Snippet: Multivariable analysis of OS in stage 0–1 patients.

Article Snippet: Following this, the slides were blocked with 10% normal mouse serum for 10 min at room temperature and incubated overnight at 4°C with the anti-IGF1R antibody (NB110-87052, 1:200; Novus Biologicals LLC, Centennial, CO, USA).

Techniques: Adjuvant