Journal: PLOS Pathogens

Article Title: EBV infection outcomes determined by monocyte and T REG -driven immune dynamics in an ex vivo pbmc model

doi: 10.1371/journal.ppat.1013746

Figure Lengend Snippet: (A) UMAP of single-cell transcriptomes from all donors and conditions, annotated by major immune cell types. (B) UMAP plots and pie charts showing relative frequencies of immune lineages across mock and EBV-infected conditions. (C) Bar plot showing the change in cell type proportions (Δ = EBV – Mock) for each donor, grouped by LCL outcome (n = 2). (D) Subset analysis of CD4 ⁺ T cells showing relative frequencies of Treg and naive CD4 ⁺ cells per donor and condition. (E) Quantification of gp350 ⁺ B cells at Day 7 post-infection in LCL-made and LCL-failed donors PBMCs treated with IgG1 isotype control (Mock), EBV + IgG1 isotype control or EBV + RG6292. Bars represent the number of gp350 ⁺ cells within the CD19 ⁺ B cell compartment (mean ± SEM; p < 0.05, ns = not significant; a paired two-sided Student’s t-test (n = 3)). (F) Representative flow cytometry plot of CD23 and CD20 expression on CD19 ⁺ B cells at Day 21 post-infection. Red and light blue represent EBV-infected cultures treated with isotype control or RG6292, respectively. Marginal histograms show marker intensity distribution. (G) gp350 surface expression on CD19 ⁺ B cells at Day 21 post-infection. Histogram overlay compares isotype- (red) and RG6292-treated (light blue) conditions.

Article Snippet: An equivalent concentration of human IgG1 isotype control (MedChemExpress, Cat# HY-P99001) was used in control conditions (Mock and EBV).

Techniques: Single Cell, Infection, Control, Flow Cytometry, Expressing, Marker

Journal: The Journal of Experimental Medicine

Article Title: HER2 drives lung fibrosis by activating a metastatic cancer signature in invasive lung fibroblasts

doi: 10.1084/jem.20220126

Figure Lengend Snippet: ERBB2 was the top upstream regulator in fibrotic fibroblasts in human and mouse lungs. (A and B) ERBB2 was the top upstream regulator of both human IPF lung fibroblasts (A) and bleomycin-induced mouse lung fibrotic fibroblasts (B). (C) Sorting strategy for normal and IPF lung fibroblasts for p-HER2 and total HER2 Western blot. (D) Immunostaining of HER2/p-HER2 with activated fibroblast (myofibroblast) marker, α-SMA, in normal and IPF human lung cryosections. IgG isotype was used as control for HER2 and p-HER2 staining. Higher magnifications of the boxed regions were provided. (E) Transcription of ERBB2 (HER2) in myofibroblasts in published scRNA-seq datasets. P value of each comparison, cell number ( n ) of each group, and average expressions of ERBB2 (purple dotted line) were included. (F) Cell proliferation rates of normal fibroblasts with HER2 overexpression or control fibroblasts were determined by EdU assays. (G) Fibroblast viability after Lapatinib treatment was determined by Calcein AM Assay ( n = 8 per group). (H) Fibroblast growth rate after treatment of Lapatinib at increasing concentration ( n = 6 per group). Three independent experiments were performed on fibroblasts from different patients (G and H). Data are the mean ± SEM. ns, not significant by two-way ANOVA (G). Scale bars, 20 μm (D).

Article Snippet: For the anti-ERBB2 antibody studies, mice were injected with IgG1 (InVivoPlus human IgG1 isotype control, Bio X Cell) or anti-HER2 (Pertuzumab, kindly provided by Genentech) twice per week, 100 μg/mouse from day 35 to 63.

Techniques: Western Blot, Immunostaining, Marker, Control, Staining, Comparison, Over Expression, Calcein AM Assay, Concentration Assay

Journal: The Journal of Experimental Medicine

Article Title: HER2 drives lung fibrosis by activating a metastatic cancer signature in invasive lung fibroblasts

doi: 10.1084/jem.20220126

Figure Lengend Snippet: Blocking HER2 signaling inhibited IPF lung fibroblast invasion and attenuated fibrosis. (A–C) Representative images (A) and index quantification (B and C) of migration and invasion of fibroblasts treated with increasing doses of Lapatinib or DMSO ( n = 3 per group). (D–F) Representative images (D) and index quantification (E and F) of migration and invasion of fibroblasts treated with Pertuzumab or IgG1 ( n = 3 per group). (G and H) Masson’s trichrome staining of collagen in lung sections (G) and hydroxyproline content in lung tissues (H) from NSG mice injected with SEMA7A high IPF fibroblasts and treated with Lapatinib, vehicle control, Pertuzumab, or IgG1 control ( n = 10 per group). Dash-boxed regions were shown at higher magnification. Three independent experiments were performed on fibroblasts from different patients (B, C, E, and F). Data are the mean ± SEM. Scale bar: 1 mm (A and D) and 500 μm (G). *, P < 0.05; **, P < 0.01; ***, P < 0.001; and ****, P < 0.0001 by one-way ANOVA (B, C, E, and F) and two-way ANOVA (H).

Article Snippet: For the anti-ERBB2 antibody studies, mice were injected with IgG1 (InVivoPlus human IgG1 isotype control, Bio X Cell) or anti-HER2 (Pertuzumab, kindly provided by Genentech) twice per week, 100 μg/mouse from day 35 to 63.

Techniques: Blocking Assay, Migration, Staining, Injection, Control

Journal: Molecular Cell

Article Title: Single-Cell Analyses Reveal Megakaryocyte-Biased Hematopoiesis in Myelofibrosis and Identify Mutant Clone-Specific Targets

doi: 10.1016/j.molcel.2020.04.008

Figure Lengend Snippet:

Article Snippet: Monoclonal Anti-DNP antibody, human IgG1 (N297A) isotype control , ACRO Biosystems , DNP-MB273.

Techniques: Recombinant, Giemsa Stain, Staining, Sample Prep, Selection, Antibody Labeling, TA Cloning, Expressing, Control, Microscopy, Sequencing, Oligo Synthesis, Software