Journal: Scientific Reports

Article Title: Cancer-Associated fibroblasts regulate the development of cholangiocarcinoma through IL-6/STAT3/AKR1C3 signaling axis

doi: 10.1038/s41598-026-37583-y

Figure Lengend Snippet: The molecular mechanism of STAT3 signaling pathway regulated by AKR1C3 in CCA. (A) After treatment with IL-6, the expression levels of proliferation, invasion, drug resistance, glycolysis, and STAT3 signaling-associated proteins were detected by Western blot in QBC939 cells. IL-6, 30 ng/ml. (B) The mRNA expression levels of the SOCSs family in QBC939 cells after AKR1C3 knockdown were detected by Q-PCR. * P < 0.05 (C) The protein expression levels of SOCS1 and SOCS3 in CCA cells after AKR1C3 knockdown and overexpression were measured by Western blot. (D) Western blot was performed to assess the ubiquitination degradation of SOCS1 in shAKR1C3- or shCtrl-transfected QBC939 cells treated with 50 µM cyclohexane (CHX) at different times. (E) Co-IP assay was conducted to investigate the protein interaction between AKR1C3 and SOCS1 in QBC939 cells.

Article Snippet: Antibodies against SOCS3 (68631 S) and PFK-1 (55028-1-AP) were obtained from Proteintech. (Hubei, China).

Techniques: Expressing, Western Blot, Knockdown, Over Expression, Ubiquitin Proteomics, Transfection, Co-Immunoprecipitation Assay

Journal: Neurotherapeutics

Article Title: Regulatory T cells attenuate astrogliosis via IL-10/STAT3/PKC/vimentin signaling and promote neurological recovery after spinal cord injury

doi: 10.1016/j.neurot.2025.e00827

Figure Lengend Snippet: IL-10 supplementation and PKC phosphorylation inhibition recapitulate Treg-mediated downregulation of vimentin in vitro . A Representative immunofluorescence images of Foxp3 (green) and IL-10 (red) of magnetically sorted Tregs from mouse spleen. B ELISA quantification of IL-10 levels in supernatants of isolated Tregs. Cohen's d = 4.7545. C Experimental timeline of in vitro IL-10 intervention in astrocytes (rIL-10: recombinant IL-10; αIL-10, IL-10 neutralizing antibody). D-H Western blot quantification of p-STAT3/STAT3 (η 2 = 0.8939), SOCS3 (η 2 = 0.9199), p -PKC/PKC (η 2 = 0.9414), and vimentin (η 2 = 0.9252) protein levels in astrocytes following IL-10 intervention. I Representative immunofluorescence images of vimentin in astrocytes following IL-10 intervention. J Experimental timeline of in vitro PKC phosphorylation modulation in astrocytes (Go6976: PKC inhibitor; PMA: PKC activator). K, L Western blot quantification of vimentin protein levels in astrocytes following PKC modulation. η 2 = 0.8597. M, N Immunofluorescence quantification of vimentin levels in astrocytes following PKC modulation. η 2 = 0.6114. Data are mean ± SEM (n = 6 independent cultures). Statistical analyses were performed using two-tailed Student's t -test (B) or one-way ANOVA (E-H, L, N). P values are indicated on the graphs. n.s., not significant.

Article Snippet: The following primary antibodies were used for Western blot assays: GAPDH (Cell Signaling Technology, 2118S), Vimentin (Abcam, AB92547), PTPσ (Solarbio, K109903P), PKCα (Proteintech, 21991-1-AP), Phospho-PKCα (Thr638) (Proteintech, 29123-1-AP), STAT3 (Cell Signaling Technology, 12640), p-STAT3 (Cell Signaling Technology, 9145), and SOCS3 (Proteintech, 66797-1-lg).

Techniques: Phospho-proteomics, Inhibition, In Vitro, Immunofluorescence, Enzyme-linked Immunosorbent Assay, Isolation, Recombinant, Western Blot, Two Tailed Test

Journal: Frontiers in immunology

Article Title: Ozone promotes macrophage efferocytosis and alleviates neuropathic pain by activating the AMPK/Gas6-MerTK/SOCS3 signaling pathway.

doi: 10.3389/fimmu.2024.1455771

Figure Lengend Snippet: FIGURE 4 Ozone suppresses neuroinflammatory responses by upregulating SOCS3 expression. (A, B) The SOCS3 and SOCS1 expression in BMDM were examined (n=3). (C-E) qPCR was performed to detect the transcription levels of inflammatory cytokines in BMDMs (n=3). Significant differences were revealed following one-way ANOVA (A–E) (*p < 0.05, **p < 0.01, and ***p < 0.001).

Article Snippet: The following reagents and antibodies were used in the study: bactin antibody from ABclonal (China), p-AMPK antibody from Cell Signaling Technology (USA), SOCS3 antibody and Protein S antibody from Santa Cruz Biotechnology (USA), Gas6 antibody from Abcam (USA), p-MerTK antibody from Thermo Fisher Scientific(USA), AICAR from Selleckchem (USA), Compound C from MedChemExpress (USA), UNC2541 from Topscience (China), BCECF from APExBIO (USA), Secondary antibodies for Western blot from Sigma (USA).

Techniques: Expressing

Journal: Frontiers in immunology

Article Title: Ozone promotes macrophage efferocytosis and alleviates neuropathic pain by activating the AMPK/Gas6-MerTK/SOCS3 signaling pathway.

doi: 10.3389/fimmu.2024.1455771

Figure Lengend Snippet: FIGURE 5 Ozone alleviates neuropathic pain by activating the AMPK/Gas6-MerTK/SOCS3 signaling pathway. (A) After establishing the mouse CCI model for 14 days, ozone (30mg, i.p.) was administered for 7 days. The sciatic nerves of mice were collected to measure the p-AMPK expression level. (B) Mice received AICAR (0.25mg/kg) and CC (100mg) pretreatment around the sciatic nerve half an hour in advance. Ozone (30mg, i.p.) was administered for 7 days, and the sciatic nerves were collected to investigate p-AMPK levels (n=6). (C, D) Western blot was used to examine the Gas6 expression of sciatic nerves and plasma in CCI mice (n=6). (E, F) After establishing the mouse CCI model for 14 days, mice received CC (100mg) pretreatment around the sciatic nerve half an hour in advance. Ozone (30mg, i.p.) was administered for 7 days, and the expression levels of Gas6 in the sciatic nerves and plasma were detected (n=6). (G, H) Western blot was used to measure the expression levels of Protein S (n=6). (I, J) p-ADAM17 and p- MerTK expression in the sciatic nerves were detected by western blot (n=6). (K, L) The expression levels of SOCS3 and SOCS1 in the sciatic nerves were examined (n=6). Significant differences were determined by one-way ANOVA (A-L). (*p < 0.05, **p < 0.01, and ***p < 0.001).

Article Snippet: The following reagents and antibodies were used in the study: bactin antibody from ABclonal (China), p-AMPK antibody from Cell Signaling Technology (USA), SOCS3 antibody and Protein S antibody from Santa Cruz Biotechnology (USA), Gas6 antibody from Abcam (USA), p-MerTK antibody from Thermo Fisher Scientific(USA), AICAR from Selleckchem (USA), Compound C from MedChemExpress (USA), UNC2541 from Topscience (China), BCECF from APExBIO (USA), Secondary antibodies for Western blot from Sigma (USA).

Techniques: Expressing, Western Blot, Clinical Proteomics

Journal: Frontiers in immunology

Article Title: Ozone promotes macrophage efferocytosis and alleviates neuropathic pain by activating the AMPK/Gas6-MerTK/SOCS3 signaling pathway.

doi: 10.3389/fimmu.2024.1455771

Figure Lengend Snippet: FIGURE 7 Schematic model indicates that ozone promotes macrophage efferocytosis and alleviates neuropathic pain. Ozone activities AMPK/GAS6/MerTK signaling pathway to promote macrophage efferocytosis and clear the apoptotic cells, inducing SOCS3- mediated neuroinflammation tolerance to attenuate neuropathic pain.

Article Snippet: The following reagents and antibodies were used in the study: bactin antibody from ABclonal (China), p-AMPK antibody from Cell Signaling Technology (USA), SOCS3 antibody and Protein S antibody from Santa Cruz Biotechnology (USA), Gas6 antibody from Abcam (USA), p-MerTK antibody from Thermo Fisher Scientific(USA), AICAR from Selleckchem (USA), Compound C from MedChemExpress (USA), UNC2541 from Topscience (China), BCECF from APExBIO (USA), Secondary antibodies for Western blot from Sigma (USA).

Techniques: