Journal: Science Advances

Article Title: Activation of the YAP1/pSTAT3/NRP1 axis in peritendinous sensory nerves promotes tendon healing

doi: 10.1126/sciadv.aec1272

Figure Lengend Snippet: ( A ) The total protein levels of YAP1, pSTAT3 Tyr 705 , and STAT3 with and without ES. β-actin was used as an internal control. n = 3. ( B ) The nuclear protein levels of YAP1 and pSTAT3 Tyr 705 with and without ES. Histone H3 was used as an internal control. n = 3. ( C ) Intersection of neurite outgrowth–related genes across multiple databases. ( D and E ) Enrichment pathways of neurite outgrowth–related genes transcribed via pSTAT3 Tyr 705 , as predicted by GO and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses, respectively. ( F ) qRT-PCR results for the mRNA expression of neurite outgrowth–related genes transcribed via pSTAT3, including MAP3K13, NRP1, PTPRF, and GSK3B. n = 3. ( G ) Protein levels of NRP1 with and without ES. β-actin was used as an internal control. n = 3. ( H ) Predicted STAT3 binding motif sequence from the JASPAR database. ( I ) Dual-luciferase assays. n = 3. ( J ) Schematic representation of the NRP1 promoter region, highlighting the predicted STAT3 binding site (blue box) and the locations of forward (F) and reverse (R) primers used for ChIP-PCR. ( K ) ChIP-PCR analysis revealed that compared with the IgG control, the NRP1 promoter fragment was enriched in the chromatin precipitated by the anti-STAT3 antibody, confirming that STAT3 occupied the predicted site. n = 3. ( L ) Expression levels of CGRP in the supernatant of DRGn with and without ES. n = 8. n.s., not significant.

Article Snippet: Culture supernatants from treated DRGn were collected, and concentrations of CGRP (lot no. E-EL-R0135, Elabscience, China) were quantified using ELISA kits according to the manufacturer’s protocols.

Techniques: Control, Quantitative RT-PCR, Expressing, Binding Assay, Sequencing, Luciferase

Journal: Science Advances

Article Title: Activation of the YAP1/pSTAT3/NRP1 axis in peritendinous sensory nerves promotes tendon healing

doi: 10.1126/sciadv.aec1272

Figure Lengend Snippet: ( A ) Schematic diagram of DRGn treatment and detection. ( B and C ) Protein expression levels and quantitative results of pSTAT3 Tyr 705 and NRP1 in vitro in the absence or presence of shSTAT3 and ES. β-actin was used as an internal control. n = 3. ( D and E ) Protein expression levels and quantitative results of YAP1 and NRP1 in vitro in the absence or presence of shYAP1 and ES. β-actin was used as an internal control. n = 3. ( F to I ) Protein expression levels and quantitative results of pSTAT3 Tyr 705 , YAP1, and NRP1 in the absence or presence of shSTAT3, shYAP1, and ES. n = 3. ( J ) Schematic illustration of the cross-talk between DRGn and HUVECs. ( K and L ) Expression levels of CGRP in the supernatant of DRGn under different treatments. n = 8. ( M ) Wound scratch assay in HUVECs under different treatments. n = 3. Ctrl, control.

Article Snippet: Culture supernatants from treated DRGn were collected, and concentrations of CGRP (lot no. E-EL-R0135, Elabscience, China) were quantified using ELISA kits according to the manufacturer’s protocols.

Techniques: Expressing, In Vitro, Control, Wound Healing Assay

Journal: Journal of Pharmaceutical Analysis

Article Title: Luteolin attenuates RA-associated chronic pain by targeting the LDHA/H3K9la/NFATC2 axis to suppress Th17 cell differentiation and central infiltration

doi: 10.1016/j.jpha.2025.101373

Figure Lengend Snippet: Luteolin (LUT) alleviates collagen-induced arthritis (CIA)-associated chronic pain by reversing central sensitization. (A) The schematic of CIA model establishment and intervention with different doses of LUT. (B) Mechanical pain threshold in each group ( n = 8 per group). (C) Thermal withdrawal latency in each group ( n = 8 per group). (D) Immunofluorescence was used to detect the expression of cFos proto-oncogene (cFos) and calcitonin gene-related peptide (CGRP) in the spinal dorsal horn (SDH) of mice across groups. Specifically, c-Fos expression was quantified by counting positive cells per mm 2 , while CGRP levels were assessed based on fluorescence intensity ( n = 4 per group). ∗ P < 0.05 and ∗∗ P < 0.01, compared with control group; # P < 0.05 and ## P < 0.01, compared with CIA model group, by repeated-measures one-way analysis of variance (ANOVA) or two-way ANOVA followed by post hoc Dunnett's multiple comparisons test. i.g. q.d.: intragastric administration once daily; DAPI: 4′,6-diamidino-2-phenylindole.

Article Snippet: : PTM-1419RM, PTM-1406RM, and PTM-1413RM; PTM Bio Inc.), rabbit anti-histone H3 (Cat. No.: ab1791; Abcam), mouse anti-CD68 (Cat. No.: sc-20060; Santa Cruz Biotechnology), rabbit anti-cFos proto-oncogene (cFos) antibody (Cat. No.: ab222699; Abcam), rabbit anti-calcitonin gene-related peptide (CGRP) antibody (Cat. No.: 14959; Cell Signaling Technology, Inc.), rabbit anti-IL-1β antibody (Cat. No.: 12242; Cell Signaling Technology, Inc.), rabbit anti-IL-6 antibody (Cat. No.: 12912; Cell Signaling Technology, Inc.), rabbit anti-TNF-α antibody (Cat. No.: 11948; Cell Signaling Technology, Inc.), rabbit anti-NFATC2 antibody (Cat. No.: 5861; Cell Signaling Technology, Inc.), rabbit anti-CD68 (Cat. No.: 97778; Cell Signaling Technology, Inc.), rabbit anti-CD40 (Cat. No.: 86165; Cell Signaling Technology, Inc.), mouse anti-NFATC2 antibody (Cat. No.: sc-7296; Santa Cruz Biotechnology), mouse anti-PRKCE antibody (Cat. No.: sc-1681; Santa Cruz Biotechnology), mouse anti-CCR6 antibody (Cat. No.: MAB590; Bio-Techne, Minneapolis, MN, USA), rabbit anti-CCL20 antibody (Cat. No.: ab9829; Abcam), rabbit anti-β-actin monoclonal antibody (Cat. No.: 4970; Cell Signaling Technology, Inc.), goat anti-rabbit IgG heavy and light chains (H&L) (Alexa Fluor® 488) (Cat. No.: ab150077; Abcam), and goat anti-mouse IgG H&L (Alexa Fluor® 647) (Cat. No.: ab150115; Abcam).

Techniques: Immunofluorescence, Expressing, Fluorescence, Control