Journal: Frontiers in Cardiovascular Medicine

Article Title: Activation of Toll-Like Receptor 9 Impairs Blood Flow Recovery After Hind-Limb Ischemia

doi: 10.3389/fcvm.2018.00144

Figure Lengend Snippet: TLR9 activation promoted TNF-α expression in macrophages. (A) Activation of TLR9 by ODN1826 (0.1-1.0 μM) for 4 h promoted the expression of TNF-α in wild-type macrophages but not in Tlr9 −/− macrophages ( N = 4, per group). (B) Wild-type or Tlr9 −/− macrophages were treated with plasma obtained from wild-type mice which received femoral ligation or sham operation for 4 h. Plasma from mice with ischemic muscle increased TNF-α expression in wild-type macrophages compared with that from sham-operated mice, however, plasma from mice with ischemic muscle did not show this response in Tlr9 −/− macrophages ( N = 4-6, per group). (C) ODN1826 promoted degradation of IκBα in wild-type macrophages in time dependent manner, suggesting the activation of NF-κB signaling. In Tlr9 −/− macrophages, this response was not observed. * P < 0.05 and *** P < 0.001. All values are mean ± SEM.

Article Snippet: Peritoneal macrophages were stimulated with ODN1826 or control-ODN1826 (GeneDesign, Inc.), synthetic oligonucleotides that contain unmethylated CpG, for indicated time.

Techniques: Activation Assay, Expressing, Ligation

Journal: Frontiers in Cardiovascular Medicine

Article Title: Activation of Toll-Like Receptor 9 Impairs Blood Flow Recovery After Hind-Limb Ischemia

doi: 10.3389/fcvm.2018.00144

Figure Lengend Snippet: TLR9-induced macrophage activation accelerated cell death of HUVEC. (A) HUVEC were treated with the CM obtained from wild-type or Tlr9 −/− macrophages treated with ODN1826 or control-ODN1826. At 72 h after treatment, the viability of HUVEC determined by MTS assay was significantly reduced by CM obtained from wild-type macrophage activated by ODN1826, although the viability of HUVEC was not affected by CM obtained from Tlr9 −/− macrophage ( N = 5, per group). (B) Stimulation with TNF-α for 24 h decreased cell viability in HUVEC as determined by MTS assay ( N = 4-6, per group). * P < 0.05 and ** P < 0.01. All values are mean ± SEM.

Article Snippet: Peritoneal macrophages were stimulated with ODN1826 or control-ODN1826 (GeneDesign, Inc.), synthetic oligonucleotides that contain unmethylated CpG, for indicated time.

Techniques: Activation Assay, MTS Assay

Journal: Frontiers in Immunology

Article Title: The Small GTPase RHOA Links SLP65 Activation to PTEN Function in Pre B Cells and Is Essential for the Generation and Survival of Normal and Malignant B Cells

doi: 10.3389/fimmu.2022.842340

Figure Lengend Snippet: Impaired humoral immune response of RhoA fl/fl Cd21 +/Cre mice. ( A ) Six-eight weeks old RhoA fl/fl (N=7) and RhoA fl/fl Cd21 +/Cre (N=8) mice were immunized with 100µg NP(24)KLH + 100µg CpG-ODN1826 in PBS or with 100µg CpG-ODN1826 (control). Serum level of NP specific IgM (A) and IgG (B) antibodies were measured by ELISA at the specified days after first immunization (day 0) and booster on day 21. No NP-specific antibodies were detected in control mice injected with adjuvant (CpG-ODN1826) hence excluded from the figure. Statistical analysis- One-way ANOVA. (C) The decline of anti-NP-IgG titers was calculated as the ratio of IgG titers at day 28 relative to day 56. Statistical significance was determined using unpaired t-test, two-tailed. *p<0.05, **p<0.01, ***p< 0,001, ****p<0.0001, ns- not significant.

Article Snippet: RhoA fl/fl (N=7) and RhoA fl/fl Cd21 +/Cre (N=8) mice of 6-8 weeks old were immunized with 100 µg NP(24)KLH (Biosearch Technologies California, N-5060-5) + 100µg CpG-ODN1826 (Biomer, custom design) in PBS or with 100µg CpG-ODN1826 (control immunization).

Techniques: Enzyme-linked Immunosorbent Assay, Injection, Two Tailed Test