Journal: eLife

Article Title: Syntaxin11 deficiency inhibits CRAC channel priming to suppress cytotoxicity and gene expression in T lymphocytes independent of membrane trafficking

doi: 10.7554/eLife.111335

Figure Lengend Snippet: ( A–B ) Measurement of total Orai1 levels in the plasma membrane of STX11-depleted cells by flow-cytometry. U2OS cells stably expressing Orai1-BBS-YFP were transduced with Scr (black) or STX11 (red) shRNA, stimulated with 1 µM TG (broken lines) or control (solid lines) and incubated with alpha-bungarotoxin alexa fluor 647 (BTX-AF647). Total Orai1-YFP levels ( A ) and BTX-AF647 binding to surface Orai1 ( B ) were measured where binding to wildtype U2OS cells was used as the background fluorescence. N=3 ( C ) Representative quantitative PCR to assess the efficiency of knockdown in shRNA-treated Orai1-BBS-YFP expressing U2OS cells from ( A–B ). Total RNA extracted from cells treated with STX11 shRNA was subjected to qPCR analysis using Taqman probes for STX11. Data were normalized to RPL30 housekeeping gene. ( D–E ) Measurement of thapsigargin (TG) induced SOCE in U2OS cells treated with STX11 RNAi in ( A–B ). ( D ) Representative averaged traces of Fura-2 calcium imaging assay performed on cells in panels ( A–B ). ( E ) Bar plot showing relative mean % SOCE ± SE from three independent experiments in ( D ). Figure 2—figure supplement 2—source data 1. Folder containing the original flow cytometry files and gating strategy for . Figure 2—figure supplement 2—source data 2. Numerical data underlying the quantitative analyses represented in .

Article Snippet: Cell line ( Homo sapiens ) , U2OS cell line , ATCC , HTB-96 (RRID: CVCL_0042 ) , .

Techniques: Clinical Proteomics, Membrane, Flow Cytometry, Stable Transfection, Expressing, Transduction, shRNA, Control, Incubation, Binding Assay, Fluorescence, Real-time Polymerase Chain Reaction, Knockdown, Analysis, Imaging

Journal: eLife

Article Title: Syntaxin11 deficiency inhibits CRAC channel priming to suppress cytotoxicity and gene expression in T lymphocytes independent of membrane trafficking

doi: 10.7554/eLife.111335

Figure Lengend Snippet:

Article Snippet: Cell line ( Homo sapiens ) , U2OS cell line , ATCC , HTB-96 (RRID: CVCL_0042 ) , .

Techniques:

Journal: International Journal of Genomics

Article Title: Integrative Single‐Cell and Bulk Transcriptomics Identifies a Proliferative Malignant State and Supports a Functional Contribution of TROAP in Osteosarcoma

doi: 10.1155/ijog/1407947

Figure Lengend Snippet: Integrated single‐cell landscape of human osteosarcoma. (A) Harmony‐integrated UMAP of 39,063 quality‐controlled cells colored by global cluster. (B) The same embedding colored by patient, showing cross‐patient mixing after integration. (C) Reviewed major‐cell‐type annotations displayed on the integrated UMAP; labels mark lineage centroids. (D) Relative abundance of major cell types in each patient. Colors are consistent between panels C and D.

Article Snippet: HOS and U2OS human osteosarcoma cell lines were obtained from the American Type Culture Collection (ATCC; Manassas, Virginia, United States; HOS, CRL‐1543; U2OS, HTB‐96).

Techniques: Single Cell

Journal: International Journal of Genomics

Article Title: Integrative Single‐Cell and Bulk Transcriptomics Identifies a Proliferative Malignant State and Supports a Functional Contribution of TROAP in Osteosarcoma

doi: 10.1155/ijog/1407947

Figure Lengend Snippet: TROAP depletion reduces clonogenic growth, overall wound closure, migration, and invasion in osteosarcoma cells. (A) Immunoblot analysis of TROAP in HOS and U2OS cells transfected with siNC, siTROAP‐1, or siTROAP‐2; beta‐actin is the loading control. (B) Colony‐formation images and colony counts. (C) Scratch‐wound images at 0 and 24 h and overall wound closure. Because the assay was not proliferation‐matched, it is not interpreted as direct migration evidence. (D) Transwell migration and invasion images and cell counts. Data are mean + /−standard deviation from three independent biological experiments. One‐way ANOVA with Dunnett′s test compared each siTROAP group with siNC; ∗∗∗ p < 0.001.

Article Snippet: HOS and U2OS human osteosarcoma cell lines were obtained from the American Type Culture Collection (ATCC; Manassas, Virginia, United States; HOS, CRL‐1543; U2OS, HTB‐96).

Techniques: Migration, Western Blot, Analysis, Transfection, Control, Colony Assay, Standard Deviation