Journal: Frontiers in Immunology

Article Title: Platelet-rich plasma-derived microRNA let-7a-5p alleviates knee osteoarthritis by regulating macrophage polarization and improving inflammatory microenvironment

doi: 10.3389/fimmu.2026.1756467

Figure Lengend Snippet: Validation of the targeting relationship between let-7a-5p and MAPK8. (A) Schematic representation of the predicted complementary binding site between let-7a-5p and the 3′-UTR of MAPK8. (B) Relative mRNA expression level of MAPK8 in macrophages overexpressing let-7a-5p detected by RT-qPCR. (C) WB analysis of MAPK8 protein expression in macrophages overexpressing let-7a-5p. (D) Inhibitory effect of let-7a-5p on MAPK8 expression was assessed via a dual luciferase reporter assay. ( **P < 0.01, ***P < 0.001).

Article Snippet: Subsequently, the tissue sections and cells were incubated overnight at 4°C with primary antibodies against iNOS (Proteintech 22226-1-AP), CD206 (Proteintech 18704-1-AP) and MAPK8 (Proteintech 66210-1-Ig).

Techniques: Biomarker Discovery, Binding Assay, Expressing, Quantitative RT-PCR, Luciferase, Reporter Assay

Journal: Frontiers in Immunology

Article Title: Platelet-rich plasma-derived microRNA let-7a-5p alleviates knee osteoarthritis by regulating macrophage polarization and improving inflammatory microenvironment

doi: 10.3389/fimmu.2026.1756467

Figure Lengend Snippet: Effect of PRP on expression of let-7a-5p and MAPK8. (A) Relative mRNA expression levels of let-7a-5p in knee joint sections of each group detected by RT-qPCR. (B) iNOS and CD206 co-immunolabeld with MAPK8 and counter-stained with DAPI in synovial tissues (Scale bar: 50 μm). (C, D) Quantification of iNOS and CD206 expression co-localized with MAPK8. ( **P < 0.01).

Article Snippet: Subsequently, the tissue sections and cells were incubated overnight at 4°C with primary antibodies against iNOS (Proteintech 22226-1-AP), CD206 (Proteintech 18704-1-AP) and MAPK8 (Proteintech 66210-1-Ig).

Techniques: Expressing, Quantitative RT-PCR, Staining

Journal: Frontiers in Immunology

Article Title: Platelet-rich plasma-derived microRNA let-7a-5p alleviates knee osteoarthritis by regulating macrophage polarization and improving inflammatory microenvironment

doi: 10.3389/fimmu.2026.1756467

Figure Lengend Snippet: The let-7a-5p/MAPK8 axis regulates macrophage polarization and inflammatory cytokine release in vitro . (A) IF staining showing expression of iNOS and CD206 in macrophages (Scale bar: 50 μm). (B) Quantification of iNOS-positive cell rate in transfected macrophages. (C) Quantification of CD206-positive cell rate in transfected macrophages. (D-G) Relative mRNA expression levels of pro-inflammatory cytokine (IL-1β and TNF-α) and anti-inflammatory cytokine (IL-4 and IL-10) in transfected macrophages detected by RT-qPCR. ( *P < 0.05, **P < 0.01, ns no significance).

Article Snippet: Subsequently, the tissue sections and cells were incubated overnight at 4°C with primary antibodies against iNOS (Proteintech 22226-1-AP), CD206 (Proteintech 18704-1-AP) and MAPK8 (Proteintech 66210-1-Ig).

Techniques: In Vitro, Staining, Expressing, Transfection, Quantitative RT-PCR

Journal: FEBS letters

Article Title: JNK1 inhibits transcriptional and pro-apoptotic activity of TAp63γ.

doi: 10.1016/j.febslet.2015.10.028

Figure Lengend Snippet: Fig. 1. JNK1 directly binds to TAp63c at its transactivation domain. (A), Hela cells transfected with Flag-JNK1 were lysed and subjected to GST pull-down assay with GST alone or GST-TAD. 10 lg of total protein from the same cell lysates were directly loaded as an input control. The adsorbates were analyzed by immunoblotting analysis (IB) using anti-Flag (top panel). Comparable amounts of GST and GST-TAD fusion proteins were shown by staining the membrane with Coomassie blue R-250 (bottom panel). (B and C) H1299 cells were co-transfected with HA-TAp63c and Flag-JNK1. Whole-cell lysates were subjected to immunoprecipitation (IP) with normal mouse IgG, anti-HA or anti-Flag. IP products were subjected to IB analysis with indicated primary antibodies and trueblot secondary antibody.

Article Snippet: JNK1 siRNA or scrambled control (Santa Cruz) was transfected with Lipofectamin 2000 to knock down endogenous JNK1.

Techniques: Transfection, Pull Down Assay, Control, Western Blot, Staining, Membrane, Immunoprecipitation

Journal: FEBS letters

Article Title: JNK1 inhibits transcriptional and pro-apoptotic activity of TAp63γ.

doi: 10.1016/j.febslet.2015.10.028

Figure Lengend Snippet: Fig. 2. Overexpression of JNK1 inhibits TAp63c transactivity. (A) H1299 cells were transfected with a mixture of Bax-Luc and TK-Renilla plus HA-TAp63c or (and) Flag-JNK1 plasmid; (B) H1299 cells were transfected with a mixture of Bax-Luc and TK-Renilla plus HA-TAp63c and increasing amounts of Flag-JNK1 plasmid. Firefly and Renilla luciferase activities were measured. The Bax-Luc activity was normalized to Renilla activity and presented as percentage of Bax-Luc expression with standard deviation (n = 3). Bax-Luc expression in cells transfected with TAp63c alone with Bax-Luc/TK-Renilla mixture was set as 100%. IB analysis was performed to detect the expression of TAp63c and JNK1 using anti-HA and anti-Flag respectively.

Article Snippet: JNK1 siRNA or scrambled control (Santa Cruz) was transfected with Lipofectamin 2000 to knock down endogenous JNK1.

Techniques: Over Expression, Transfection, Plasmid Preparation, Luciferase, Activity Assay, Expressing, Standard Deviation

Journal: FEBS letters

Article Title: JNK1 inhibits transcriptional and pro-apoptotic activity of TAp63γ.

doi: 10.1016/j.febslet.2015.10.028

Figure Lengend Snippet: Fig. 3. Ser12 in TAp63c is critical for JNK1-mediated trans-inhibition. H1299 cells were transfected with Bax-Luc/TK-Renilla mixture plus HA-TAp63c (wild-type or S12A mutant) or (and) Flag-JNK1 plasmid. Firefly and Renilla luciferase activities were measured. The Bax-Luc activity was normalized to Renilla activity and presented as percentage of Bax-Luc expression with standard deviation (n = 3). Bax-Luc expression in cells transfected with wild-type TAp63c alone with Bax-Luc/ TK-Renilla mixture was set as 100 percent. IB analysis was performed to detect the expression of TAp63c and JNK1 using anti-HA and anti-Flag respectively.

Article Snippet: JNK1 siRNA or scrambled control (Santa Cruz) was transfected with Lipofectamin 2000 to knock down endogenous JNK1.

Techniques: Inhibition, Transfection, Mutagenesis, Plasmid Preparation, Luciferase, Activity Assay, Expressing, Standard Deviation

Journal: FEBS letters

Article Title: JNK1 inhibits transcriptional and pro-apoptotic activity of TAp63γ.

doi: 10.1016/j.febslet.2015.10.028

Figure Lengend Snippet: Fig. 4. Inhibition or knockdown of JNK1 increases TAp63c transactivity. (A) H1299 cells were transfected with Bax-Luc/TK-Renilla mixture with or without HA-TAp63c, and treated with JNK inhibitor SP600125 (JNKin); (B) H1299 cells were transfected with Bax-Luc/TK-Renilla mixture plus HA-TAp63c or (and) siJNK1. Firefly and Renilla luciferase activities were measured. The Bax-Luc activity was normalized to Renilla activity and presented as percentage of Bax-Luc expression with standard deviation (n = 3). Bax-Luc expression in cells transfected with TAp63c alone with Bax-Luc/TK-Renilla mixture was set as 100%. IB analysis was performed to detect the expression of TAp63c, total or phosphorylated (p-JNK1) JNK1 using anti-HA, antibody specific to total or T183-phosphorylated JNK1, respectively.

Article Snippet: JNK1 siRNA or scrambled control (Santa Cruz) was transfected with Lipofectamin 2000 to knock down endogenous JNK1.

Techniques: Inhibition, Knockdown, Transfection, Luciferase, Activity Assay, Expressing, Standard Deviation

Journal: FEBS letters

Article Title: JNK1 inhibits transcriptional and pro-apoptotic activity of TAp63γ.

doi: 10.1016/j.febslet.2015.10.028

Figure Lengend Snippet: Fig. 5. JNK1 inhibits TAp63c-mediated cell apoptosis. H1299 cells were transfected with HA-TAp63c and (or) Flag-JNK1. 48 h post transfection, cells were trypsinized and subjected to IB (A) or flow cytometry analysis (B) of apoptosis. Cleaved PARP1 (CL-PARP), JNK1 and TAp63c were detected with antibody to N-terminal cleaved PARP1, Flag, or HA respectively.

Article Snippet: JNK1 siRNA or scrambled control (Santa Cruz) was transfected with Lipofectamin 2000 to knock down endogenous JNK1.

Techniques: Transfection, Cytometry

Journal: Frontiers in pharmacology

Article Title: α-Mangostin Alleviated Inflammation in Rats With Adjuvant-Induced Arthritis by Disrupting Adipocytes-Mediated Metabolism-Immune Feedback.

doi: 10.3389/fphar.2021.692806

Figure Lengend Snippet: FIGURE 8 | Effects of MAN on inflammatory macrophages co-cultured with pre-adipocytes. (A) levels of IL-1β and IL-6 in medium from macrophages culture chamber; (B) expression of mRNA IL-1β and iNOS in LPS-primed macrophages co-cultured with pre-adipocytes in the presence of MAN or not; (C) expression of protein TLR4, p65, p-p65, JNK, p-INK and COX-2 in above macrophages; (D) quantification result of assay (C). Statistical significance: *p < 0.05 and **p < 0.01 compared with LPS-stimulated macrophages; #p < 0.05 and ##p < 0.01 compared with LPS-primed macrophages in the presence of untreated pre-adipocytes (n 3).

Article Snippet: The primary anti-SIRT1 (A17307), PPAR-c (A19676), p-JNK (AP0473), JNK (A11119), p-p65 (AP0123), p65 (A11201), NAMPT (A0256), toll like receptor 4 (TLR4, A5258), cyclooxygenase-2 (COX-2, A1253), stearoyl-CoA desaturase 1 (SCD-1, A16429), CYP4A1 (A19662) and β-actin (AC026) antibodies were procured from AB clonal Technology (Wuhan, China).

Techniques: Cell Culture, Expressing

Journal: Frontiers in pharmacology

Article Title: α-Mangostin Alleviated Inflammation in Rats With Adjuvant-Induced Arthritis by Disrupting Adipocytes-Mediated Metabolism-Immune Feedback.

doi: 10.3389/fphar.2021.692806

Figure Lengend Snippet: FIGURE 9 | Pre-adipocytes culture medium affected the status of macrophages. (A) levels of IL-1β and IL-6 and MCP-1 released by macrophages receiving pre- adipocytes culture medium treatment; (B) expression of protein TLR4, p65, p-p65, JNK, p-INK and COX-2 in macrophages obtained from assay (A); (C) quantification result of assay (B). Statistical significance: *p < 0.05 and **p < 0.01 compared with macrophages cultured with medium from untreated pre-adipocytes; #p < 0.05 and ##p < 0.01 compared with macrophages cultured with medium from LPS-primed pre-adipocytes [n 4 in image (A), n 3 in image (C)].

Article Snippet: The primary anti-SIRT1 (A17307), PPAR-c (A19676), p-JNK (AP0473), JNK (A11119), p-p65 (AP0123), p65 (A11201), NAMPT (A0256), toll like receptor 4 (TLR4, A5258), cyclooxygenase-2 (COX-2, A1253), stearoyl-CoA desaturase 1 (SCD-1, A16429), CYP4A1 (A19662) and β-actin (AC026) antibodies were procured from AB clonal Technology (Wuhan, China).

Techniques: Expressing, Cell Culture