Journal: PLoS ONE

Article Title: IGFBP2 promotes immunosuppression associated with its mesenchymal induction and FcγRIIB phosphorylation in glioblastoma

doi: 10.1371/journal.pone.0222999

Figure Lengend Snippet: (A-C) The mouse splenocytes (SPCs) were co-cultured with GL261 cells for 6 days and then collected to analyze the number of CD8 + and CD4 + T cells by fluorescence activated cell sorting (FACS) under the setting of IGFBP2KD (A), anti-IGFBP2 (B) or overexpressed IGFBP2 and its mutant (C). The experiments were repeated at least three times. Representative IHC images of CD8 + T cells (D-E), CD163 + M2 macrophages (G-H), and pY513-CD19 + cells (J-K) in tumor-bearing brains of mice treated with anti-IGFBP2 or IgG. 100X (left), 400X (right). N = 3. (F, I) The percentage of tumor infiltrating CD8 + T cells (F) and CD163 + M2 macrophages in F4/80 + macrophages (I) analyzed by FACS after anti-IGFBP2 or IgG treatment. N = 5. The data are mean ± SD. A statistical significance was calculated by an unpaired Welch’s t test. * P<0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: For anti-IGFBP2 antibody treatment, 2.0 μg/ml of a rabbit anti-IGFBP2 monoclonal antibody (anti-IGFBP2mAb) (bs-1108R, Bioss) or isotype IgG control (bs-0295P, Bioss) was added to GL261 culture for 3 days and collected for western blotting.

Techniques: Cell Culture, Fluorescence, FACS, Mutagenesis

Journal: PLoS ONE

Article Title: IGFBP2 promotes immunosuppression associated with its mesenchymal induction and FcγRIIB phosphorylation in glioblastoma

doi: 10.1371/journal.pone.0222999

Figure Lengend Snippet: ( A) An association of the enrichment score of the MES signature with FcγRIIB expression analyzed by using the TCGA-human glioma database. R = Pearson’s correlation coefficient. (B) Representative IHC images of IGFBP2 protein and phosphorylated FcγRIIB (p-FcγRIIB) levels in human GBMs (hGBMs) or normal brains stained in tissue microarrays. (C) An association of IGFBP2 protein and p-FcγRIIB levels in human GBMs. R, the Pearson correlation coefficient. (D-I), The percentage of p-FcγRIIB + cells in the SPCs after co-cultured with GL261 cells treated with sh1RNA-IGFBP2 (D-E), anti-IGFBP2 (F-G) or overexpressing IGFBP2 or IGFBP2mt (H-I) analyzed by FACS. (J-K), Representative IHC images of p-FcγRIIB staining in tumor-bearing brains treated with anti-IGFBP2 or IgG. N = 3. (L) The percentage of p-FcγRIIB + cells in infiltrating immune cells in GL261 tumors analyzed by FACS. N = 5. The data are mean ± SD. A statistical significance was computed by an unpaired Welch’s t test. * P < 0.05, ** p < 0.01.

Article Snippet: For anti-IGFBP2 antibody treatment, 2.0 μg/ml of a rabbit anti-IGFBP2 monoclonal antibody (anti-IGFBP2mAb) (bs-1108R, Bioss) or isotype IgG control (bs-0295P, Bioss) was added to GL261 culture for 3 days and collected for western blotting.

Techniques: Expressing, Staining, Cell Culture

Journal: PLoS ONE

Article Title: IGFBP2 promotes immunosuppression associated with its mesenchymal induction and FcγRIIB phosphorylation in glioblastoma

doi: 10.1371/journal.pone.0222999

Figure Lengend Snippet: (A-B) The time course induction of FcγRIIB phosphorylation on CD19 + B cells in SPCs co-cultured with IGFBP2OE GL261 or control cells. The percentage of CD19 L p-FcγRIIB H , CD19 L p-FcγRIIB - , and CD19 H p-FcγRIIB L B cell subsets was analyzed by FACS. L is low, H is high. (C-H) The percentage of p-FcγRIIB + cells in CD19 + B cells after co-cultured with GL261 cells treated with shRNA (C-D), anti-IGFBP2 (E-F) or overexpressing IGFBP2 or its mutant (G-H). (I-J) The percentage of CD19 + p-FcγRIIB + B cells in infiltrating immune cells (I) and p-FcγRIIB + cells in CD19 + B cells in anti-IGFBP2 or IgG tumors analyzed by FACS(J). (K) The ratio of CD19 + B cells versus CD19 + p-FcγRIIB + B cells. (L-M) The percentage of F4/80 + p-FcγRIIB + macrophages in infiltrating immune cells (L) and p-FcγRIIB + cells in F4/80 + macrophages (M) in anti-IGFBP2 or IgG tumors analyzed by FACS. N = 5. The data are mean ± SD. A statistical significance was calculated by an unpaired Welch's t test. * P < 0.05, ** P < 0.01.

Article Snippet: For anti-IGFBP2 antibody treatment, 2.0 μg/ml of a rabbit anti-IGFBP2 monoclonal antibody (anti-IGFBP2mAb) (bs-1108R, Bioss) or isotype IgG control (bs-0295P, Bioss) was added to GL261 culture for 3 days and collected for western blotting.

Techniques: Cell Culture, shRNA, Mutagenesis

Journal: PLoS ONE

Article Title: IGFBP2 promotes immunosuppression associated with its mesenchymal induction and FcγRIIB phosphorylation in glioblastoma

doi: 10.1371/journal.pone.0222999

Figure Lengend Snippet: ( A) Representative T2-weighted MRI of GL261-bearing brains of mice treated with anti-IGFBP2 or IgG after 3 and 5 weeks of GL261cell injection. (B) The tumor growth curve in anti-IGFBP2 and IgG groups. (C) The Kaplan-Meier survival plot of GL261-bearing mice treated with anti-IGFBP2 or IgG. Log-rank test was used to compare the difference of the median survival time between the two groups. N = 5.

Article Snippet: For anti-IGFBP2 antibody treatment, 2.0 μg/ml of a rabbit anti-IGFBP2 monoclonal antibody (anti-IGFBP2mAb) (bs-1108R, Bioss) or isotype IgG control (bs-0295P, Bioss) was added to GL261 culture for 3 days and collected for western blotting.

Techniques: Injection

Journal: Blood cells, molecules & diseases

Article Title: CXCL15/Lungkine has Suppressive Activity on Proliferation and Expansion of Multi-potential, Erythroid, Granulocyte and Macrophage Progenitors in S-Phase Specific Manner

doi: 10.1016/j.bcmd.2021.102594

Figure Lengend Snippet: Expression of CXCL15 by phenotyped HSC and HPC subsets in normal mouse BM. A) Expression of Cxcl15 mRNA as analyzed by microarray for selected primitive and mature mouse hematopoietic cells was obtained from the BloodSpot database (28–30). Data shown is for microarray probe 1456428_at. LMPP=lymphoid primed multipotent progenitors; CLP = common lymphoid progenitor; MkE = megakaryocyte erythroid progenitor. B-D) Wildtype mouse BM was harvested, cells were stained for cell surface markers to define specific subpopulations of HSC/HPC, and cells were permeabilized and stained with FITC-anti-CXCL15 antibody. B) Representative FACS plot demonstrating CXCL15 protein expression in Lin+ or LSK cells from murine BM. Isotype control is shown. C) Percentage of cells in the indicated subpopulations that are positive for CXCL15 protein expression using an isotype control to gate CXCL15+ versus CXCL15- cells. D) Calculated mean fluorescence intensities for FITC (conjugated to anti-CXCL15) for the indicated subpopulations of cells. Example of Flow analysis shown in Suppl. Fig. 1.

Article Snippet: Cells were then fixed and permeabilized with Cytofix/Cytoperm (BD Biosciences), then stained with anti-CXCL15-FITC antibody (Bioss Antibodies bs-2554R-FITC) or isotype-FITC control (Bioss Antibodies bs-0295P-FITC).

Techniques: Expressing, Microarray, Staining, Fluorescence