Journal: Cell

Article Title: B cell-specific XIST complex enforces X-inactivation and restrains atypical B cells

doi: 10.1016/j.cell.2021.02.015

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Electroporated cells were immediately transferred to pre-warmed B cell expansion medium (RPMI 1640 +10% FBS +1XGlutaMAX (Thermo Fisher Scientific) + 50 ng/mL IL2 + CellXVivo Human B cell expander (1:250 dilution, R&D system)).

Techniques: Recombinant, Sequencing, Mass Spectrometry, Software

Journal: Cell

Article Title: B cell-specific XIST complex enforces X-inactivation and restrains atypical B cells

doi: 10.1016/j.cell.2021.02.015

Figure Lengend Snippet: A. Bar plot showing significantly enriched biological pathway and diseases from genes that are upregulated after XIST perturbation in B cell line. B. GSEA analysis showing enrichment of XIST-dependent and -independent X-linked genes in PBMC of female SLE patients compared to healthy donors. Normalized Enrichment Score (NES) and FDR q value are shown. C-D. Uniform manifold approximation and projection (UMAP) plot showing B cell subsets from female COVID-19 patients (C), and XIST escape score (D). E. Diffusion map showing the pseudotime trajectory using naive cells as root (left). B cell subset projected on diffusion map (right). F. Violin plot showing the distribution of XIST escape score between CD11c+ atypical memory B cells and conventional memory B cells in female (left) and male (right) COVID-19 patients. P value was from unpaired t-test. G-H. UMAP showing XIST escape score of CD11c+ atypical memory cells and conventional memory B cells in kidney of female lupus patients (G), and in joint tissue of female RA patients (H). Violin plot showing the distribution of XIST escape score in two B cell subsets (right). P value was from unpaired t-test. I. Schematic view of XIST KO in human primary B cells upon B cell activation by BCR crosslinking and TLR7 ligand stimulation. J. Flow cytometry analysis of viable CD19+ human primary B cells between AAVS1 control and XIST KO group(left). Box plot showing the frequency of IgD+CD11c-, IgD-CD11c+, IgDCD11c-B cells among total B cells in two to three replicates from 8 female donors. K. Flow cytometry analysis of IgG+ B cells gated on IgD-CD11c+ B cell subset. Box plot showing the frequency of IgD-IgG+CD11c+ and IgD-IgG-CD11c+ B cells among total B cells. P value in J-K was from paired t-test. L. A proposed model of XCI maintenance in human B cells. See also Figure S7 and Table S7

Article Snippet: Isolated B cells were cultured in IMDM medium supplemented with 10% FBS and 55 mM beta-mercaptoethanol at 1X10 6 cell/mL and primed with CellXVivo Human B cell expander (1:250 dilution, R&D system), 50 ng/mL IL10 (Cat#200-10-2ug, PeproTech), 10 ng/mL IL15 (Cat#200-15-2ug, PeproTech), 50 ng/mL IL2 (Cat#200-02-10ug, PeproTech) for two days.

Techniques: Diffusion-based Assay, Activation Assay, Flow Cytometry, Control

Journal: Cell

Article Title: B cell-specific XIST complex enforces X-inactivation and restrains atypical B cells

doi: 10.1016/j.cell.2021.02.015

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Isolated B cells were cultured in IMDM medium supplemented with 10% FBS and 55 mM beta-mercaptoethanol at 1X10 6 cell/mL and primed with CellXVivo Human B cell expander (1:250 dilution, R&D system), 50 ng/mL IL10 (Cat#200-10-2ug, PeproTech), 10 ng/mL IL15 (Cat#200-15-2ug, PeproTech), 50 ng/mL IL2 (Cat#200-02-10ug, PeproTech) for two days.

Techniques: Virus, Recombinant, Sequencing, Mass Spectrometry, Software