Journal: Blood Advances

Article Title: Clinical characteristics, management, and hematopoietic cell transplantation of patients with TLR8 gain-of-function

doi: 10.1182/bloodadvances.2025016338

Figure Lengend Snippet: TLR8 variants associated with disease. (A) Schematic representation of TLR8 protein structure showing the localization of TLR8 variants associated with disease, including 2 published variants resulting in disease phenotypes other than INFLTR8 (p.G572V results in TLR8 protein loss, dysregulation of TLR7/8 signaling, and anemia and autoinflammation and p.V434L in inherited anemia ). (B) NF-κB reporter cells (HEK-Blue Null1 cells) that do not express endogenous TLR8 were transfected with WT TLR8, patient TLR8 variants (encoding p.P432L, p.P432Q, p.P432R), or a loss-of-function TLR8 variant (encoding p.D543A) and stimulated with the indicated doses of the TLR8-specific agonist TL8-506. The newly identified patient TLR8 variants lead to GOF in TLR8 activity as measured by NF-κB transcriptional activity. Data are represented as mean ± standard deviation (SD) of biological replicates and representative of 2 independent experiments, each performed in duplicate. ∗∗∗ P < .001, by 2-way analysis of variance (ANOVA) test. (C) VAF in the peripheral blood by age of disease onset for patients with mosaic variants in TLR8 . (D) Number of patients with the most commonly reported disease manifestations. LOF, loss-of-function; SEAP, secreted alkaline phosphatase.

Article Snippet: New variants identified in patients 7 and 8 (p.P432Q and p.P432R) or reported in a family with anemia (p.V434L) were tested using HEK-Blue Null1 NF-κB reporter cells (InvivoGen) lacking TLR8 expression as previously described.

Techniques: Transfection, Variant Assay, Activity Assay, Standard Deviation

Journal: Journal of Inflammation Research

Article Title: Mechanism of BMSCs Inhibiting Microglia Activation and Neuroinflammatory Response to Alleviate BCP by Regulating TLR4/ NF-κB Signaling Pathway

doi: 10.2147/JIR.S553485

Figure Lengend Snippet: BMSCs significantly inhibited the expression of TLR4/NF-κB signaling pathway related markers in BCP. ( A and B ) The relative expression level of TLR4, p-NF-κB p65, and NF-kB p65 proteins were determined by Western blotting. ( C and D ) Co-localization staining of Iba-1 and p-NF-κB p65/TLR4 in spinal dorsal horn, and the red box indicates the region magnified on the right. Mean ± SD, one-way ANOVA , * p <0.05, ** p <0.01.

Article Snippet: The following antibodies were used: Iba-1 (Abcam, ab178846, 1:1000), TLR4 (Santa Cruz Biotechnology, sc-293072, 1:1000), NF-κB p65 (Bioss, bs-0465R, 1:1000), p-NF-κB p65 (Affinity, AF2006, 1:1000), and GAPDH (HUABIO, R1210-1, 1:5000).

Techniques: Expressing, Western Blot, Staining

Journal: Journal of Inflammation Research

Article Title: Mechanism of BMSCs Inhibiting Microglia Activation and Neuroinflammatory Response to Alleviate BCP by Regulating TLR4/ NF-κB Signaling Pathway

doi: 10.2147/JIR.S553485

Figure Lengend Snippet: The effect of BMSCs on TLR4/NF-κB signaling pathway in BV2 cells. ( A and B ) The relative expression levels of TLR4, p-NF-κB p65, NF-κB p65 in different groups by Western blotting. ( C and D ) Immunostaining of p-NF-κB p65 and TLR4 in different groups. Mean ± SD, one-way ANOVA , * p <0.05, ** p <0.01.

Article Snippet: The following antibodies were used: Iba-1 (Abcam, ab178846, 1:1000), TLR4 (Santa Cruz Biotechnology, sc-293072, 1:1000), NF-κB p65 (Bioss, bs-0465R, 1:1000), p-NF-κB p65 (Affinity, AF2006, 1:1000), and GAPDH (HUABIO, R1210-1, 1:5000).

Techniques: Expressing, Western Blot, Immunostaining