Journal: International journal of molecular sciences

Article Title: Targeting TLR2/Rac1/cdc42/JNK Pathway to Reveal That Ruxolitinib Promotes Thrombocytopoiesis.

doi: 10.3390/ijms232416137

Figure Lengend Snippet: Figure 8. Biophysical validation reveals TLR2 as a new protein target of ruxolitinib. (A) Venn di‑ agram shows the common targets of Ruxolitinib and thrombocytopenia. The intersecting part rep‑ resents the common targets between Ruxolitinib and thrombocytopenia; (B) PPI network for identi‑ fying core targets of Ruxolitinib against thrombocytopenia through the screening conditions of De‑ gree > 47, BC > 0.002858932, CC > 0.507867733; (C) TLR2 and ligands (ruxolitinib) by molecular dock‑ ing; (D). Representative immunoblot images and biochemical quantification of TLR2 after treatment with Ruxolitinib (5, 10, and 20 µM) in Meg‑01 cells for 5 day (E) The DARTS assay for target valida‑ tion. TLR2 protein stability was increased upon Ruxolitinib (200 µM) treatment in Meg‑01 lysates. Pronase was added using several dilutions (1:500, 1:1000, or 1500) from 50 µg/mL stock for 10 min at 40 ◦C; (F) The DARTS assay demonstrated the dose‑dependent binding of Ruxolitinib to TLR2. Treatment with pronase (1:1000) was conducted for 10 min at 40 ◦C; (G) Meg‑01 cells were treated with ruxolitinib (20 µM), C29 (50 µM), ruxolitinib (20 µM) + C29 (50 µM) for 5 days. FCM analysis of the expression of CD41 and CD42b. (H) The histogram shows the percentage of CD41+/CD42b+

Article Snippet: The primary antibodies were as follows: FOS (Proteintech, USA, 66590‐1‐lg), EGR1 (Proteintech, USA, 22008–1‐AP), RUNX1 (Proteintech, USA, 25315‐1‐AP), TLR2 (Pro‐ teintech, USA, 17236‐1‐AP), Rac1/cdc42 (CST, USA, 4651S), JNK (Abmart, China, T55490), p‐JNK (Abmart, China, T55541), NF‐E2 (Proteintech, USA, 11089‐1‐AP), and GAPDH (Pro‐ teintech, USA, 60004‐1‐lg).

Techniques: Biomarker Discovery, Western Blot, Binding Assay, Expressing

Journal: International journal of molecular sciences

Article Title: Targeting TLR2/Rac1/cdc42/JNK Pathway to Reveal That Ruxolitinib Promotes Thrombocytopoiesis.

doi: 10.3390/ijms232416137

Figure Lengend Snippet: Figure 10. Schematic illustration of the role of ruxolitinib in MK differentiation and platelet pro‑duction. Ruxolitinib induces the expression of various cytokines and TLR2, activates the Rac1/cdc42/JNK signaling pathway, and leads to the expression of AP‑1, EGR1, RUNX1, and NF‑E2. As a result, the activation of AP‑1, EGR1, RUNX1, NF‑E2 promote the expression of genes related to MK differentiation and thrombopoiesis. These genes contribute to MK maturation and platelet for‑ mation and promote the recovery of bone marrow and spleen MKs and accelerate platelet production in RI‑mice. PPF: proplatelet‑forming MK.

Article Snippet: The primary antibodies were as follows: FOS (Proteintech, USA, 66590‐1‐lg), EGR1 (Proteintech, USA, 22008–1‐AP), RUNX1 (Proteintech, USA, 25315‐1‐AP), TLR2 (Pro‐ teintech, USA, 17236‐1‐AP), Rac1/cdc42 (CST, USA, 4651S), JNK (Abmart, China, T55490), p‐JNK (Abmart, China, T55541), NF‐E2 (Proteintech, USA, 11089‐1‐AP), and GAPDH (Pro‐ teintech, USA, 60004‐1‐lg).

Techniques: Expressing, Activation Assay

Journal: BMC Musculoskeletal Disorders

Article Title: TLR2 and IRF8 are the biomarkers associated with hypoxia in steroid-induced osteonecrosis of the femoral head via bioinformatics analysis and experimental validation

doi: 10.1186/s12891-025-09132-7

Figure Lengend Snippet: ROC curves analysis of 5 hub genes. ( A ) ROC curve of TLR2 in SONFH. ( B ) ROC curve of IRF8 in SONFH. ( C ) ROC curve of CXCL8 in SONFH. ( D ) ROC curve of CD86 in SONFH. ( E ) ROC curve of MAPK1 in SONFH.

Article Snippet: Human TLR2 ELISA kit (No. CSB-E11781h, Cusabio, Wuhan, China), human IRF8 ELISA kit (No. A- QEK00354 , Biogradetech, USA), and human CXCL8 ELISA kit (No. CSB-E04641h, Cusabio, Wuhan, China), human CD86 ELISA kit (No. CSB-E08542h, Cusabio, Wuhan, China), and human MAPK1 ELISA kit (No. A- QEK07595 , Biogradetech, USA) were used for ELISA according to the manufacturer’s instructions.

Techniques:

Journal: Frontiers in Pharmacology

Article Title: Integrating RNA-sequencing and network analysis to explore the mechanism of topical Pien Tze Huang treatment on diabetic wounds

doi: 10.3389/fphar.2023.1288406

Figure Lengend Snippet: Tlr9, klrk1, Nod2, Tlr2, and Ifng were identified as vital targets. (A) A network of upregulated DEGs of PZH. (B) A network of downregulated DEGs of PZH. (C) The GO terms of upregulated DEGs. (D) The GO terms of downregulated DEGs. (E) Venn diagram of DEGs of PZH, BBR and NR1. (F) GO analysis between the PZH group and BBR group. (G) GO analysis between the PZH group and NR1 group. (H) A network of vital biological processes and potential vital targets involved in three biological processes were labeled with a red frame. (I) A network of potential vital targets of PZH. (J) Venn diagram of downregulated DEGs and unique DEGs of PZH.

Article Snippet: We evaluated the expression of transforming growth factor-β1 (TGF-β1) (E-EL-0162c, Elabscience), matrix metallopeptidase 9 (MMP9) (E-EL-R3021, Elabscience), interleukin-1β (IL-1β) (E-EL-R0012c, Elabscience), toll-like receptor 2 (TLR2) (E-EL-R0907c, Elabscience), Interferon Gamma (IFN-γ) (E-EL-R0009c, Elabscience) in rat skin tissue by ELISA.

Techniques: Labeling

Journal: Frontiers in Pharmacology

Article Title: Integrating RNA-sequencing and network analysis to explore the mechanism of topical Pien Tze Huang treatment on diabetic wounds

doi: 10.3389/fphar.2023.1288406

Figure Lengend Snippet: PZH significantly decreased the expression of NKG2-D, IFN-γ, NOD2, and TLR2, and inhibited NF-κB signaling pathway in diabetic wounds. (A) IF staining and its quantification results for NKG2-D (red). Scale bar: 50 μm ( n = 3). (B) IF staining and its quantification results for NOD2 (red). Scale bar: 50 μm ( n = 3). (C) IF staining and its quantification results for TLR2 (green). Scale bar: 50 μm ( n = 3). (D) IF staining and its quantification results for p65 (green). Scale bar: 50 μm ( n = 3). (E) Western blotting for NKG2-D, IκBα and RELB. (F–H) Quantification results of Western blotting of NKG2-D, IκBα and RELB ( n = 3). (I,J) TLR2 and IFN-γ ELISA results ( n = 6). (K) The network of active ingredients-drug targets-PZH-regulated targets. Data are presented as the mean ± SD, # p < 0.05, ## p < 0.01, ### p < 0.001, #### p < 0.0001 vs. control, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 vs. hfdSTZ.

Article Snippet: We evaluated the expression of transforming growth factor-β1 (TGF-β1) (E-EL-0162c, Elabscience), matrix metallopeptidase 9 (MMP9) (E-EL-R3021, Elabscience), interleukin-1β (IL-1β) (E-EL-R0012c, Elabscience), toll-like receptor 2 (TLR2) (E-EL-R0907c, Elabscience), Interferon Gamma (IFN-γ) (E-EL-R0009c, Elabscience) in rat skin tissue by ELISA.

Techniques: Expressing, Staining, Western Blot, Enzyme-linked Immunosorbent Assay, Control