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: The interfering RNA sequences used for IGFBP2 knockdown.

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: Sequencing

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 the expression of IGFBP2 using the TCGA-gliomas database. R = Pearson’s correlation coefficient. (B-C) Effect of IGFBP2 knockdown with shRNA (IGFBP2KD) (B) or an anti-IGFBP2 antibody (anti-IGFBP2) (C) on mesenchymal marker protein levels in mouse GBM GL261 cells. (D) Effect of overexpressed IGFBP2 wildtype (IGFBP2OE) or mutant (IGFBP2mt) with the change of RGD to RGE on mesenchymal marker protein levels. The experiments were repeated at least three times. A statistical significance was calculated using 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: Expressing, shRNA, Marker, 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-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: P values in Fig 4C–4H .

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:

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: Molecular Therapy. Methods & Clinical Development

Article Title: Enzyme Replacement Therapy Can Reverse Pathogenic Cascade in Pompe Disease

doi: 10.1016/j.omtm.2020.05.026

Figure Lengend Snippet: AT-GAA Reversed the Levels of the Autophagic Markers and Alleviated the Burden of Accumulated Protein Aggregates and ER-Stress in Muscle from KO Mice Muscle biopsies were collected from age and sex-matched WT, untreated (KO), and AT-GAA-treated KO (KO-ERT) mice. (A) Western blot analysis of muscle lysates from WT, untreated KO (KO), and treated KO (KO-ERT) mice with the indicated antibodies (n = 4 for each group; n = 5 for western blot with LC3 antibody). (B) Western blot analysis of muscle lysates from WT, untreated KO (KO), and treated KO (KO-ERT) mice with K63-linked ubiquitin specific antibody (n = 4 for each group). Western blot with anti-GAPDH and Ponceau S staining were used as loading controls. (C) Western blot analysis of muscle lysates from WT, untreated KO (KO), and treated KO (KO-ERT) mice with anti-Grp 78 antibody (ER-stress marker; n = 4 for each group); GAPDH was used as loading control. Each data point represents an individual mouse. Statistical significance was determined by one-way ANOVA. Graphs represent mean ± SD. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ∗∗∗∗p < 0.0001.

Article Snippet: SQSTM1/p62 (#ab56416; mouse monoclonal), galectin 9 (#ab69630; rabbit polyclonal), Ubiquitin K48 (Linkage Specific) (#ab140601; rabbit monoclonal), and glyceraldehyde-3-phosphate dehydrogenase (#ab9485; rabbit polyclonal) were purchased from Abcam.

Techniques: Western Blot, Ubiquitin Proteomics, Staining, Marker, Control

Journal: Cell

Article Title: Macrophages direct location-dependent recall of B cell memory to vaccination.

doi: 10.1016/j.cell.2025.04.005

Figure Lengend Snippet: Figure 4. SSMs in the dLN subcapsular niche control Bmem cell fate (A) Schematic for comparing swapping the source of T cell help in Bmem recall response. (B) Representative flow cytometric analysis of boosted dLN (left) and ndLN (right) showing effect of swapping T cell help from OT-II mTfh to Smarta naive T cells. (C) Enumeration of donor-derived antigen-specific total B cells, GC B cells, and PCs in boosted dLN and ndLN in recall response to HEL-OVA and HEL-gp61 boost. Data were pooled from three in- dependent experiments. (D) Schematic for determining the impact of SSM depletion on Bmem recall responses in dLN vs. ndLN. (E) Representative plots showing the effect of isotype control (left) or anti-CSF1R mAb (right) on GC B cell and PC differentiation in boosted dLN (top) and boosted ndLN (bottom). (F) Enumeration of donor-derived antigen-specific total B cells, GC B cells, and PCs in boosted dLN and ndLN treated with isotype control or anti-CSF1R. Data were pooled from two independent experiments. In (C) and (F), each circle or square represents one mouse. Line represents median.

Article Snippet: In vivo monoclonal antibody treatment Anti-mouse CSF1R mAb (clone AFS98, Bio X Cell) was used to deplete SSMs.

Techniques: Control, Derivative Assay

Journal: Cell

Article Title: Macrophages direct location-dependent recall of B cell memory to vaccination.

doi: 10.1016/j.cell.2025.04.005

Figure Lengend Snippet: Figure 6. Primed SSMs mediate superior Bmem responses agnostic of the antigen (A) Schematic for synchronous priming of the contralateral ndLN with the irrelevant antigen R-phycoerythrin (PE). (B) Enumeration of total number of donor-derived antigen-specific B cells, GC B cells, and PCs in contralaterally HEL-OVA-boosted ndLN treated with isotype control or anti-CSF1R mAb. (C) Percentage of GC B cells and PCs within donor-derived antigen-specific B cells in contralaterally HEL-OVA-boosted ndLN treated with isotype control or anti- CSF1R mAb. Data were pooled from two independent experiments. Each circle represents one mouse. Line and bar represent median.

Article Snippet: In vivo monoclonal antibody treatment Anti-mouse CSF1R mAb (clone AFS98, Bio X Cell) was used to deplete SSMs.

Techniques: Derivative Assay, Control

Journal: Molecular cancer therapeutics

Article Title: DHA-SBT-1214 Taxoid Nanoemulsion and Anti-PD-L1 Antibody Combination Therapy Enhances Anti-Tumor Efficacy in a Syngeneic Pancreatic Adenocarcinoma Model

doi: 10.1158/1535-7163.MCT-18-1046

Figure Lengend Snippet: In vivo efficacy of the PD-L1 antibody in combination to different therapeutic drugs including gemcitabine solution, Abraxane™, paclitaxel in solution, DHA-SBT-1214 in solution and NE-DHA-SBT-1214 against Panc02 induced syngeneic mice tumors. (A)– Graph summarizing all treatment modalities. The values are means ± SD (n=3). Significant differences are indicated as follows: *p < 0.05, and **p< 0.01. (B)– Tumor images taken at the time of harvest from different treatment modalities. (3b-A)– Tumors from mice treated with vehicle; (3b-B)– Three tumors each from PD-L1 (200μg) treated mice; (3b-C, D)–Tumors from Abraxane™ plus IgG or PD-L1 (200μg) treated mice respectively; (3b-E)– Tumors from NE-DHA-SBT-1214 (10mg/kg) plus IgG (200μg) treated mice; (3b-F, G)– Tumors from gemcitabine plus IgG or PD-L1 (200μg) treated mice respectively; (3b-H)– Tumors from NE-DHA-SBT-1214 (10mg/kg) plus PD-L1 (200μg) treated mice; (3b-I, J)– Tumors from NE-DHA-SBT-1214 (25mg/kg) plus IgG or PD-L1 (200μg) treated mice respectively. (C)– Graph for all the tumors from (3B) to show their progression over time.

Article Snippet: In Vivo Single and Combination Therapies Mouse antibody against PD-L1 (10F.9G2) and relevant isotype IgG control was purchased from Bio X Cell.

Techniques: In Vivo

Journal: Molecular cancer therapeutics

Article Title: DHA-SBT-1214 Taxoid Nanoemulsion and Anti-PD-L1 Antibody Combination Therapy Enhances Anti-Tumor Efficacy in a Syngeneic Pancreatic Adenocarcinoma Model

doi: 10.1158/1535-7163.MCT-18-1046

Figure Lengend Snippet: In vivo PD-L1 surface protein expression in response to different therapeutic modalities. (A)– mRNA expression of PD-L1 from different mouse tumor treatment groups analyzed using RT-PCR. Relative gene expression for RT-PCR data was calculated relative to murine β-actin. (B)– Tumor tissue lysate from different treated groups was prepared and protein level of different proteins was analyzed using western blotting. (C)– The bands corresponding to PD-L1 were quantified using Image J software and was normalized relative to band intensities for the corresponding Histone 3 loading controls. The bar represents the mean ± standard deviation of data from at least 3 independent experiments; *p<0.05, **p<0.01.

Article Snippet: In Vivo Single and Combination Therapies Mouse antibody against PD-L1 (10F.9G2) and relevant isotype IgG control was purchased from Bio X Cell.

Techniques: In Vivo, Expressing, Reverse Transcription Polymerase Chain Reaction, Gene Expression, Western Blot, Software, Standard Deviation

Journal: Molecular cancer therapeutics

Article Title: DHA-SBT-1214 Taxoid Nanoemulsion and Anti-PD-L1 Antibody Combination Therapy Enhances Anti-Tumor Efficacy in a Syngeneic Pancreatic Adenocarcinoma Model

doi: 10.1158/1535-7163.MCT-18-1046

Figure Lengend Snippet: Histopathological evaluation of the Panc02-induced tumor tissues collected from control and different combination treated mice (hematoxylin & eosin staining). Significant reduction in tumor stroma observed with combination of NE-DHA-SBT-1214 and anti-PD-L1 treated groups. The images were taken at 63x magnification.

Article Snippet: In Vivo Single and Combination Therapies Mouse antibody against PD-L1 (10F.9G2) and relevant isotype IgG control was purchased from Bio X Cell.

Techniques: Control, Staining