Journal: Arthritis Research & Therapy

Article Title: Expansion of myeloid-derived suppressor cells contributes to metabolic osteoarthritis through subchondral bone remodeling

doi: 10.1186/s13075-021-02663-z

Figure Lengend Snippet: High-fat diet increases monocytic-MDSC expansion during PTOA. A Gating strategy used to define the M-MDSC subpopulation (CD11b + Ly6C + Ly6G − cells) in mouse bone marrow. Following the initial FS/SC discrimination, the gate was set on CD11b + cells. After exclusion of doublets (not shown), live CD11b + cells were gated and Ly6C + and Ly6G + populations. B Cytospin images of M-MDSC cells from the bone marrow from LFD and HFD mice (at 16 weeks). Scale bar denotes 10 microns. C HFD increases the BM M-MDSC population in mice ( n = 4 in each group) by total number and percentage with concomitant suppression of CD8 + and CD4 + T-cells compared to LFD mice ( n = 6 in each group, at 16 weeks). D HFD increases the BM M-MDSC population in DMM mice ( n = 3 in each group). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 by two-way ANOVA Tukey’s multiple comparison test

Article Snippet: The following anti-human antibodies were purchased from Miltenyi Biotec (Germany): CD11b-APC (clone M1/70, # 130-113-793), HLA-DR–PE (clone AC122, #130-113-402), CD14-fluorescein isothiocyanate (FITC) (clone Tuk4, #130-113-146), CD15-APCvio770 (clone VIMC6, #130-104-992), CD33-viobright515 (clone REA775, #130-111-027), CD66b-PEvio770 (cloneREA306, #130-119-808), and their corresponding isotype controls, along with corresponding isotype controls.

Techniques: Comparison

Journal: Arthritis Research & Therapy

Article Title: Expansion of myeloid-derived suppressor cells contributes to metabolic osteoarthritis through subchondral bone remodeling

doi: 10.1186/s13075-021-02663-z

Figure Lengend Snippet: MDSCs are expanded in obese patients with post-traumatic osteoarthritis. A Demographic of the patient population. B Gating strategy used to define the M-MDSC subpopulation (CD11b + HLADR − CD14 + CD15 − cells). The M-MDSC population is expanded in C peripheral blood and D synovial fluid of obese ( BMI > 30; N = 10) compared to non-obese ( BMI < 30; N = 5) patients diagnosed with post-traumatic osteoarthritis. Two-tailed unpaired Student t -test

Article Snippet: The following anti-human antibodies were purchased from Miltenyi Biotec (Germany): CD11b-APC (clone M1/70, # 130-113-793), HLA-DR–PE (clone AC122, #130-113-402), CD14-fluorescein isothiocyanate (FITC) (clone Tuk4, #130-113-146), CD15-APCvio770 (clone VIMC6, #130-104-992), CD33-viobright515 (clone REA775, #130-111-027), CD66b-PEvio770 (cloneREA306, #130-119-808), and their corresponding isotype controls, along with corresponding isotype controls.

Techniques: Two Tailed Test

Journal: Oncology Reports

Article Title: Adipocytes induce the resistance of ovarian cancer to carboplatin through ANGPTL4

doi: 10.3892/or.2020.7647

Figure Lengend Snippet: Downstream effect of ANGPTL4 following binding to integrin α5β1 on surface of the tumor cell membrane. (A) Co-IP was performed and the immunoprecipitates were analyzed using western blot analysis with antibodies against integrin α5β1, claudin-5, ANGPTL4 and GAPDH. (B) Statistical results of the co-IP assay. The data are presented as the mean ± SD. **P<0.01 vs. SKOV3. (C and D) The regulatory effect following silencing of integrin α5β1 on the ANGPTL4-induced resistance of SKOV3 cells in vitro . The data are presented as the mean ± SD. **P<0.01 vs. SKOV3 shMock /CM/carboplatin or SKOV3 shMock /adipocyte/carboplatin). (E and F) The regulatory effect following silencing of claudin 5 on the ANGPTL4-induced resistance of SKOV3 cells in vitro . The data are presented as the mean ± SD. **P<0.01 vs. SKOV3 shMock /CM/carboplatin or SKOV3 shMock /adipocyte/carboplatin). ANGPTL4, angiopoietin-like 4; Co-IP, co-iummunoprecipitation; sh, short hairpin; CM, conditioned medium.

Article Snippet: Subsequently, the membranes were blocked with 5% skimmed milk, prepared with the TBS-Tween-20 buffer, and incubated with primary antibody against ANGPTL4 (1:1,000; cat. no. ab2920; Abcam), integrin α5β1 (1:1,000; cat. no. orb13515; Biorbyt Ltd.), claudin 5 (1:2,000; cat. no. ab131259; Abcam), VE-cadherin (1:1,000; cat. no. 2158; Cell Signaling Technology, Inc.), Bcl-xl (1:1,000; cat. no. ab32370; Abcam), ABCB1 (1:1,000; cat. no. 13342; Cell Signaling Technology, Inc.), ABCC1 (1:1,000; cat. no. 72202; Cell Signaling Technology, Inc.), ABCG2 (1:1,000; cat. no. ab207732; Abcam), β-actin (1:10,000; cat. no. BS6007M; Bioworld Technology, Inc.), PI3K (1:2,000; cat. no. 4257; Cell Signaling Technology, Inc.), phospho-PI3K (1:1,000; cat. no. 4228; Cell Signaling Technology, Inc.), c-Myc (1:2,000; cat. no. 18583; Cell Signaling Technology, Inc.), phospho-c-Myc (1:1,000; cat. no. 13748; Cell Signaling Technology, Inc.), p65 (1:1,000; cat. no. 8242; Cell Signaling Technology, Inc.), phospho-p65 (1:1,000; cat. no. ab194726; Abcam), Akt (1:2,000; cat. no. 4691; Cell Signaling Technology, Inc.), phospho-Akt (1:2,000; cat. no. 4060; Cell Signaling Technology, Inc.) overnight at 4°C, whilst avoiding non-specific background.

Techniques: Binding Assay, Membrane, Co-Immunoprecipitation Assay, Western Blot, In Vitro

Journal: Cell Communication and Signaling : CCS

Article Title: Neutrophil membrane-coated circular RNA nanoparticles for targeted immunotherapy in HER2-positive breast cancer brain metastasis

doi: 10.1186/s12964-025-02321-w

Figure Lengend Snippet: Construction and Characterization of HER2-Targeted dHL-60 Cell Membranes. (A) Schematic representation of the lentiviral vector design used to overexpress the scFv targeting HER2. The vector includes the anti-HER2 scFv, a membrane localization signal, a FLAG tag for detection, a hinge region for flexibility, and a transmembrane domain to anchor the scFv to the cell membrane. (B) SPR analysis illustrating the binding affinity of scFv to the HER2 protein. (C) Flow cytometry analysis comparing the binding of scFv (anti-HER2) to SKBR3 cells versus a control scFv. (D) Flow cytometry assessment of CD11b expression levels in HL-60 cells before and after differentiation induction with DMSO. (E) Western blot results demonstrating the expression of adhesion molecules CD11b, CD18, CD49d, and CD31 in differentiated HL-60 cells. (F) Western blot analysis was performed using an anti-FLAG antibody to detect the FLAG-tagged scFv (anti-HER2). A band corresponding to the scFv-FLAG fusion protein is observed in dHL-60 cells transduced with the lentiviral vector, but not in control cells. (G) Flow cytometry analysis (anti-FLAG antibody) demonstrating the surface expression of scFv (anti-HER2) on dHL-60 cells overexpressing scFv (anti-HER2). (H) Illustration depicting the construction and extraction process of the cell membrane from HL-60 cells, outlining the steps involved in preparing membrane-coated nanoparticles

Article Snippet: CoraLite ® Plus 488, PE-conjugated Goat Anti-Rabbit IgG, antibodies against CD11B, CD49d, CD31, CD18, β-actin, His Tag, FLAG Tag and CXCL9 were obtained from Proteintech (Wuhan, China).

Techniques: Plasmid Preparation, Membrane, FLAG-tag, Binding Assay, Flow Cytometry, Control, Expressing, Western Blot, Transduction, Extraction