tak242  (MedChemExpress)

Journal: Neural Regeneration Research

Article Title: Porcine decellularized nerve matrix hydrogel attenuates neuroinflammation after peripheral nerve injury by inhibiting the TLR4/MyD88/NF-κB axis

doi: 10.4103/NRR.NRR-D-24-00302

Figure Lengend Snippet: TLR4/MyD88/NF-κB pathway plays an indispensable role in regulating pDNM-gel-mediated macrophage polarization and anti-inflammatory reactions in vitro . (A) WB detection of TLR4, MyD88, p-NF-κB, NF-κB, p-IκBα, and IκBα in RAW264.7 cells treated with LPS (100 ng/mL) + pDNM-gel (0.5%) for 24 hours, followed by the addition of CRX-527 (500 µM), TAK242 (100 nM), CL075 (2 µM), or T6167923 (100 µM) for another 24 hours. (B–E) Quantitative analysis of the expression levels of these four proteins from A. (F, G) Co-staining of CD68 (red) with iNOS (green) or Arg1 (green) in the LPS + pDNM-gel, LPS + pDNM-gel + CRX-527, LPS + pDNM-gel + TAK242, LPS + pDNM-gel + CL075, and LPS + pDNM-gel + T6167923 treatment groups. Nuclei were stained with DAPI (blue). Scale bars: 10 μm. (H, I) Quantification of the fluorescence intensity of iNOS and Arg1 from F and G. (J, K) Flow cytometry detection of M1-type and M2-type macrophages in each group. (L, M) Quantitative analysis of the percentages of double-positive CD68 + CD86 + and CD68 + CD206 + cells from J and K. (N, O) qRT-PCR analysis of the mRNA expression of pro-inflammatory signature genes TNF-α , IL-1 β, and IL-6 , and anti-inflammatory signature genes IL-4 , IL-10 , and TGF- β, in each group. (P, Q) ELISAs of inflammatory cytokine levels in RAW264.7 cell culture supernatants in each group. Data are expressed as mean ± SEM from three independent assays in vitro ; * P < 0.05, ** P < 0.01, *** P < 0.001. Arg1: Arginase 1; CD206: cluster of differentiation protein 206; CD68: cluster of differentiation protein 68; CD86: cluster of differentiation protein 86; DAPI: 4′,6-Diamidino-2-phenylindole; ELISA: Enzyme-linked immunosorbent assay; IL-10: interleukin-10; IL-1β: interleukin-1β; IL-4: interleukin-4; IL-6: interleukin-6; iNOS: inducible nitric oxide synthase; IκBα: nuclear factor kappa B inhibitory protein alpha; LPS: lipopolysaccharides; MyD88: myeloid differentiation factor 88; NF-κB: nuclear factor-κB; n.s,: not significant; pDNM-gel: porcine decellularized nerve matrix hydrogel; p-IκBα: phosphorylated nuclear factor kappa B inhibitory protein alpha; p-NF-κB: phosphorylated nuclear factor-κB; qRT-PCR: quantitative reverse transcription-polymerase chain reaction; TGF-β: transforming growth factor-beta; TLR4: Toll-like receptor 4; TNF-α: tumor necrosis factor-alpha.

Article Snippet: Then, the medium was added with one of the following agonists/inhibitors and treated the cells for a further 24 hours: 500 μM CRX-527 (InvivoGen, Toulouse, France, Cat# tlrl-crx527), 100 nM TAK242 (MedChemExpress, Monmouth Junction, NJ, USA, Cat# HY-11109), 2 μM CL075 (MedChemExpress, Cat# HY-117066), or 100 μM T6167923 (MedChemExpress, Cat# HY-19744; Additional Figure 3 ).

Techniques: In Vitro, Expressing, Staining, Fluorescence, Flow Cytometry, Quantitative RT-PCR, Cell Culture, Enzyme-linked Immunosorbent Assay, Reverse Transcription, Polymerase Chain Reaction

Journal: Redox Biology

Article Title: Amino acid restriction sensitizes lung cancer cells to ferroptosis via GCN2-dependent activation of the integrated stress response

doi: 10.1016/j.redox.2025.103988

Figure Lengend Snippet: F12 medium sensitizes lung cancer cells to iron-dependent lipid peroxidation . ( A, B ) Quantification by flow cytometry of oxidized BODIPY-C11 fluorescence in A549 (A) or H838 (B) cells that were cultured in RPMI or F12 medium and treated with 100 μM BSO or vehicle for 24 h. ( C ) Dose response curves for F12-cultured A549 and H838 cells treated with BSO in combination with 5 μM liproxstatin-1 (LIP-1), 5 μM ferrostatin-1 (FER-1) or 50 μM α-tocopherol (α-toco) for 72 h. ( D ) Dose response curves for F12-cultured A549 or H838 cells treated with BSO in combination with 5 μM deferoxamine (DFO) for A549 cells and 9 μM for H838 cells for 72 h. ( E ) Dose response curves for F12-cultured A549 or H838 cells treated with BSO in combination with 5 μM necrostatin-1 (NEC-1) or 10 μM ZVAD-FMK (ZVAD) for 72 h. ( F ) Dose response curves for F12-cultured A549 or H838 cells treated with BSO in combination with 4 μg/mL certolizumab (CER), 15 μM CU-CPT4a (C4a), 3 μM resatorvid (RST), or 50 μM necrostatin-1 (NEC-1) for 72 h (ND, not done). Dose response curves were normalized against the mean of the untreated samples for each condition. n = 3 replicates for all datapoints, error bars show SEM. ∗∗∗∗P < 0.0001.

Article Snippet: Drugs used were auranofin (A6733; Sigma-Aldrich), α-tocopherol (T3251; Sigma-Aldrich), bafilomycin A1 (SML1661; Sigma-Aldrich), certolizumab pegol (HYP9953; MedChemExpress), chloroquine (C6628; Sigma-Aldrich), CU-CPT4A (HY-108473; MedChemExpress), deferoxamine (D9533; Sigma-Aldrich), erastin (e7781; Sigma Aldrich), FCCP (Carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone) (HY-100410; MedChemExpress), ferrostatin-1 (SML0583; Sigma-Aldrich), l -buthionine-sulfoximine (b2515; Sigma-Aldrich), mito-TEMPO (HY–W001187; MedChemExpress), necrostatin-1 (N9037; Sigma-Aldrich), liproxstatin-1 (SML1414; Sigma-Aldrich), oligomycin (HY–N6782; MedChemExpress), puromycin dihydrochloride (A1113803; Gibco), rotenone (HY–B1756; MedChemExpress), RSL3 (SML2234; Sigma-Aldrich), resatorvid (HY-11109; MedChemExpress), and Z-VAD-FMK (V116; Sigma-Aldrich).

Techniques: Flow Cytometry, Fluorescence, Cell Culture

Journal: The Veterinary Quarterly

Article Title: An underlying mechanism of bovine mastitis: PGE 2 regulates Staphylococcus aureus -induced inflammatory response through TLR2, TLR4, and NLRP3 in macrophages

doi: 10.1080/01652176.2026.2615759

Figure Lengend Snippet: Pathogen perspective: Lipoproteins are essential for the S. aureus -induced inflammatory response in bBMMs. The bBMMs were infected with S. aureus SA113 (WT), isogenic mutant lgt ::ermB (Δ lgt ), or its complemented strain, lgt ::ermB + pRB lgt (+pRB), at MOI 10:1 or not infected. The secretion of TNF-α, IL-1β and IL-10 were detected by ELISA (6, 9, and 12 h after infection) (A–C). The activation of the MAPK (P-ERK and P-p38) and NF-κB (P-p65) pathways was evaluated by western blotting at 15, 30, and 60 min post-infection (D). TLR2 , TLR4 and NLRP3 mRNA expression levels were detected by qRT-PCR and normalized to those of the housekeeping gene β-actin at 4 h post-infection (E–G). Results are expressed as mean ± SD of three independent experiments and were analyzed using two-way ANOVA with Bonferroni’s post-hoc test. * P < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001 indicated statistically significant differences between two experimental groups. These data are representative of three independent experiments.

Article Snippet: The bBMMs (5 × 10 6 cells/well) were treated with the TLR2 inhibitor C29 (10 −5 M, MCE, Monmouth Junction, NJ, USA) for 1 h before infection; the TLR4 inhibitor TAK242 (10 −5 M, MCE, Monmouth Junction, NJ, USA) for 1 h before infection; the NLRP3 inhibitor MCC950 (10 −5 M, Cayman Chemical Company, Ann Arbor, MI, USA) for 4 h before infection; the COX-2 inhibitor CAY10404 (10 −5 M, Cayman Chemical Company, Ann Arbor, MI) was given for 40 min before infection, and the mPGES-1 inhibitor CAY10526 (10 −5 M, Cayman Chemical Company, Ann Arbor, MI) was administered for 12 h before infection; the PGE 2 (10 −5 M, Cayman Chemical Company, Ann Arbor, MI, USA) for 24 h before infection.

Techniques: Infection, Mutagenesis, Enzyme-linked Immunosorbent Assay, Activation Assay, Western Blot, Expressing, Quantitative RT-PCR

Journal: The Veterinary Quarterly

Article Title: An underlying mechanism of bovine mastitis: PGE 2 regulates Staphylococcus aureus -induced inflammatory response through TLR2, TLR4, and NLRP3 in macrophages

doi: 10.1080/01652176.2026.2615759

Figure Lengend Snippet: Host perspective: TLR2, TLR4, and NLRP3 play essential roles in the inflammatory response induced by S. aureus infection in bBMMs. bBMMs were pretreated with the TLR2 inhibitor (C29, 10 −5 M, before infection for 1 h), TLR4 inhibitor (TAK242, 10 −5 M, before infection for 1 h), and NLRP3 inhibitor (MCC950, 10 −5 M, before infection for 4 h). Then, bBMMs were infected SA113 (MOI 10:1), or uninfected. The activation of the MAPK (P-ERK and P-p38) and NF-κB (P-p65) pathways was evaluated by western blotting at 15, 30, and 60 min post-infection (A). The secretion of TNF-α, IL-1β and IL-10 were detected by ELISA (6, 9, and 12 h after infection) (B–D). Results are expressed as mean ± SD of three independent experiments and were analyzed using two-way ANOVA with Bonferroni’s post-hoc test. * p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001 indicated statistically significant differences between two experimental groups. These data are representative of three independent experiments.

Article Snippet: The bBMMs (5 × 10 6 cells/well) were treated with the TLR2 inhibitor C29 (10 −5 M, MCE, Monmouth Junction, NJ, USA) for 1 h before infection; the TLR4 inhibitor TAK242 (10 −5 M, MCE, Monmouth Junction, NJ, USA) for 1 h before infection; the NLRP3 inhibitor MCC950 (10 −5 M, Cayman Chemical Company, Ann Arbor, MI, USA) for 4 h before infection; the COX-2 inhibitor CAY10404 (10 −5 M, Cayman Chemical Company, Ann Arbor, MI) was given for 40 min before infection, and the mPGES-1 inhibitor CAY10526 (10 −5 M, Cayman Chemical Company, Ann Arbor, MI) was administered for 12 h before infection; the PGE 2 (10 −5 M, Cayman Chemical Company, Ann Arbor, MI, USA) for 24 h before infection.

Techniques: Infection, Activation Assay, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: The Veterinary Quarterly

Article Title: An underlying mechanism of bovine mastitis: PGE 2 regulates Staphylococcus aureus -induced inflammatory response through TLR2, TLR4, and NLRP3 in macrophages

doi: 10.1080/01652176.2026.2615759

Figure Lengend Snippet: Host perspective: TLR2, TLR4, and NLRP3 play essential roles in PGE 2 production induced by S. aureus infection in bBMMs. bBMMs were pretreated with the TLR2 inhibitor (C29, 10 −5 M, before infection for 1 h), TLR4 inhibitor (TAK242, 10 −5 M, before infection for 1 h), and NLRP3 inhibitor (MCC950, 10 −5 M, before infection for 4 h). Then, bBMMs were infected SA113 (MOI 10:1), or uninfected. The experession of the COX-2 and mPGES-1 was evaluated by western blotting at 12 h and 24 h post-infection (A). COX-2 and mPGES-1 mRNA expression levels were detected by qRT-PCR and normalized to those of the housekeeping gene β-actin at the 4 h and 8 h post-infection (B, C). The secretion of PGE 2 were detected by ELISA (9 h after infection) (D). Results are expressed as mean ± SD of three independent experiments and were analyzed using two-way ANOVA with Bonferroni’s post-hoc test. * p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001 indicated statistically significant differences between two experimental groups. These data are representative of three independent experiments.

Article Snippet: The bBMMs (5 × 10 6 cells/well) were treated with the TLR2 inhibitor C29 (10 −5 M, MCE, Monmouth Junction, NJ, USA) for 1 h before infection; the TLR4 inhibitor TAK242 (10 −5 M, MCE, Monmouth Junction, NJ, USA) for 1 h before infection; the NLRP3 inhibitor MCC950 (10 −5 M, Cayman Chemical Company, Ann Arbor, MI, USA) for 4 h before infection; the COX-2 inhibitor CAY10404 (10 −5 M, Cayman Chemical Company, Ann Arbor, MI) was given for 40 min before infection, and the mPGES-1 inhibitor CAY10526 (10 −5 M, Cayman Chemical Company, Ann Arbor, MI) was administered for 12 h before infection; the PGE 2 (10 −5 M, Cayman Chemical Company, Ann Arbor, MI, USA) for 24 h before infection.

Techniques: Infection, Western Blot, Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: The Veterinary Quarterly

Article Title: An underlying mechanism of bovine mastitis: PGE 2 regulates Staphylococcus aureus -induced inflammatory response through TLR2, TLR4, and NLRP3 in macrophages

doi: 10.1080/01652176.2026.2615759

Figure Lengend Snippet: Cross-talk: PGE 2 regulates TLR2, TLR4, and NLRP3 expression and inflammatory responses in S. aureus -infected bBMMs. bBMMs were pretreated with the COX-2 inhibitor ( CAY10404 , 10 −5 M, before infection for 40 min), mPGES-1 inhibitor (CAY10526, 10 −5 M, before infection for 12 h). Then, bBMMs were infected SA113 (MOI 10:1), or uninfected. The secretion of PGE 2 were detected by ELISA (9 h after infection) (A). TLR2 , TLR4 and NLRP3 mRNA expression levels were detected by qRT-PCR and normalized to those of the housekeeping gene β-actin at 4 h post-infection (B–D). The secretion of TNF-α, IL-1β and IL-10 were detected by ELISA (9 h and 12 h after infection) (E–J). Phagocytosis of Hoechst 33258 (blue)-labelled SA113 S. aureus within DiI-labelled macrophages (Orange) was analyzed by microscopy assay (×400, K). Results are expressed as mean ± SD of three independent experiments and were analyzed using two-way ANOVA with Bonferroni’s post-hoc test. * p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001 indicated statistically significant differences between two experimental groups. These data are representative of three independent experiments.

Article Snippet: The bBMMs (5 × 10 6 cells/well) were treated with the TLR2 inhibitor C29 (10 −5 M, MCE, Monmouth Junction, NJ, USA) for 1 h before infection; the TLR4 inhibitor TAK242 (10 −5 M, MCE, Monmouth Junction, NJ, USA) for 1 h before infection; the NLRP3 inhibitor MCC950 (10 −5 M, Cayman Chemical Company, Ann Arbor, MI, USA) for 4 h before infection; the COX-2 inhibitor CAY10404 (10 −5 M, Cayman Chemical Company, Ann Arbor, MI) was given for 40 min before infection, and the mPGES-1 inhibitor CAY10526 (10 −5 M, Cayman Chemical Company, Ann Arbor, MI) was administered for 12 h before infection; the PGE 2 (10 −5 M, Cayman Chemical Company, Ann Arbor, MI, USA) for 24 h before infection.

Techniques: Expressing, Infection, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Microscopy

Journal: The Veterinary Quarterly

Article Title: An underlying mechanism of bovine mastitis: PGE 2 regulates Staphylococcus aureus -induced inflammatory response through TLR2, TLR4, and NLRP3 in macrophages

doi: 10.1080/01652176.2026.2615759

Figure Lengend Snippet: Cross-talk: Excess PGE 2 exacerbates inflammation and impairs intracellular killing in S. aureus -infected bBMMs. bBMMs were pretreated with the PGE 2 (10 −6 M, before infection for 24 h). Then, bBMMs were infected SA113 (MOI 10:1), or uninfected. TLR2 , TLR4 and NLRP3 mRNA expression levels were detected by qRT-PCR and normalized to those of the housekeeping gene β-actin at 4 h post-infection (A–C). The activation of the MAPK (P-ERK and P-p38) and NF-κB (P-p65) pathways was evaluated by western blotting at 15, 30, and 60 min post-infection (D). The secretion of TNF-α, IL-1β and IL-10 were detected by ELISA (9 h and 12 h after infection) (E–G). Phagocytosis of Hoechst 33258 (blue)-labelled SA113 S. aureus within DiI-labelled macrophages (Orange) was analyzed by microscopy assay (×400, H). Results are expressed as mean ± SD of three independent experiments and were analyzed using two-way ANOVA with Bonferroni’s post-hoc test. *p < 0.05, ** p < 0.01, *** p < 0.001 and *** *p < 0.0001 indicated statistically significant differences between two experimental groups. These data are representative of three independent experiments.

Article Snippet: The bBMMs (5 × 10 6 cells/well) were treated with the TLR2 inhibitor C29 (10 −5 M, MCE, Monmouth Junction, NJ, USA) for 1 h before infection; the TLR4 inhibitor TAK242 (10 −5 M, MCE, Monmouth Junction, NJ, USA) for 1 h before infection; the NLRP3 inhibitor MCC950 (10 −5 M, Cayman Chemical Company, Ann Arbor, MI, USA) for 4 h before infection; the COX-2 inhibitor CAY10404 (10 −5 M, Cayman Chemical Company, Ann Arbor, MI) was given for 40 min before infection, and the mPGES-1 inhibitor CAY10526 (10 −5 M, Cayman Chemical Company, Ann Arbor, MI) was administered for 12 h before infection; the PGE 2 (10 −5 M, Cayman Chemical Company, Ann Arbor, MI, USA) for 24 h before infection.

Techniques: Infection, Expressing, Quantitative RT-PCR, Activation Assay, Western Blot, Enzyme-linked Immunosorbent Assay, Microscopy

Journal: The Veterinary Quarterly

Article Title: An underlying mechanism of bovine mastitis: PGE 2 regulates Staphylococcus aureus -induced inflammatory response through TLR2, TLR4, and NLRP3 in macrophages

doi: 10.1080/01652176.2026.2615759

Figure Lengend Snippet: Graphical abstract of the present study: the involvement of TLR2-, TLR4-, and NLRP3-dependent PGE 2 signaling in macrophage responses to S. aureus , which modulates inflammatory signaling and phagocytic activity and thereby contributes to the pathogenesis of bovine mastitis.

Article Snippet: The bBMMs (5 × 10 6 cells/well) were treated with the TLR2 inhibitor C29 (10 −5 M, MCE, Monmouth Junction, NJ, USA) for 1 h before infection; the TLR4 inhibitor TAK242 (10 −5 M, MCE, Monmouth Junction, NJ, USA) for 1 h before infection; the NLRP3 inhibitor MCC950 (10 −5 M, Cayman Chemical Company, Ann Arbor, MI, USA) for 4 h before infection; the COX-2 inhibitor CAY10404 (10 −5 M, Cayman Chemical Company, Ann Arbor, MI) was given for 40 min before infection, and the mPGES-1 inhibitor CAY10526 (10 −5 M, Cayman Chemical Company, Ann Arbor, MI) was administered for 12 h before infection; the PGE 2 (10 −5 M, Cayman Chemical Company, Ann Arbor, MI, USA) for 24 h before infection.

Techniques: Activity Assay

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Cathelicidin-Ka, the first frog-derived TLR2 and TLR4 agonist, induces macrophage activation and promotes inflammation

doi: 10.1007/s00018-025-06068-y

Figure Lengend Snippet: Cath-Ka directly binds to both TLR2 and TLR4. ( A ) Flow cytometry measurement of the binding reaction of FITC-labeled Cath-Ka (1.25–20 µM) with RAW264.7 cells. ( B ) The effects of TLR inhibitors on Cath-Ka-induced TNF-α production. RAW264.7 cells were pre-treated with the indicated inhibitors for 30 min and then co-cultured with Cath-Ka (10 µM) for 24 h before TNF-α production was measured by ELISA. ( C ) The effects of Cath-Ka (10 µM) on LTA (10 µg/mL) - or LPS (100 ng/mL)- induced TNF-α production in RAW264.7 cells. ( D ) Representative SPRi sensograms of Cath-Ka binding to TLR2 or TLR4 immobilized on a gold chip. Recombinant human TLR2 and TLR4 (1 mg/mL) were immobilized on an activated bare gold SPRi chip before Cath-Ka (0.25–4 µM) was used as the mobile phase. ( E ) Representative WB images of TLR2 protein pulled down by Cath-Ka. TLR2 protein of RAW264.7 cell lysates was captured with biotin-tagged Cath-Ka-bound Streptavidin resin before it was eluted and used for WB analyses. Cath-Ka-1 was used as the negative control. ( F ) Representative WB images of CETSA. RAW264.7 cells were stimulated with Cath-Ka (10 µM) or PBS for 1 h under the indicated temperatures before TLR2 (a) and TLR4 (b) protein expressions were detected by WB. Data are expressed as mean ± SD ( n = 3). *** P < 0.001 is thought statistically significant to the control group, while # P < 0.05, ## P < 0.01, and ### P < 0.001 are deemed to have significant differences to the corresponding Cath-Ka/LTA/LPS treatment groups; ns means not significant

Article Snippet: The inhibitors against TLR2 (C29, #HY-100461), TLR3 (CU-CPT 4a, #HY-108473), TLR4 (TAK-242, #HY-11109), TLR5 (TH1020, #HY-116961), TLR7/9 (Hydroxychloroquine, #HY-W031727), TLR7/8 (Enpatoran, #HY-134581), ERK (U0126, # HY-12031 A), p38 (SB203580H, # Y-10256), and JNK (SP600125, #HY-12041) were purchased from MedChem Express (Shanghai, China).

Techniques: Flow Cytometry, Binding Assay, Labeling, Cell Culture, Enzyme-linked Immunosorbent Assay, Recombinant, Negative Control, Control

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Cathelicidin-Ka, the first frog-derived TLR2 and TLR4 agonist, induces macrophage activation and promotes inflammation

doi: 10.1007/s00018-025-06068-y

Figure Lengend Snippet: Cath-Ka activates TLR2/4-MyD88-MAPKs signaling pathways in vitro and vivo. ( A , B ) Representative WB images (left) and quantitative analysis (right) of the expression of TLR2/4-MyD88-MAPKs signaling proteins activated by Cath-Ka. ( A ) RAW264.7 cells were stimulated with Cath-Ka (10 µM) for the indicated time (0.5, 1, 2, 4, and 6 h); ( B ) RAW264.7 cells were stimulated with the indicated concentrations of Cath-Ka (2.5, 5, 10, and 20 µM) for 4 h (for the detection of TLR2, TLR4, and MyD88) or 0.5 h (for the detection of MAPKs) before the proteins related to TLR2/4-MyD88-MAPKs signaling pathways were measured by WB. ( C , D ) Representative WB images (left) and quantitative analysis (right) of the effects of C29 and TAK-242 on the expression of proteins related to TLR2/4-MyD88-MAPKs signaling pathways in the presence of Cath-Ka. RAW264.7 cells were pre-treated with 10 µM C29 ( C ) or 10 µM TAK-242 ( D ) for 30 min and then were cultured with medium containing Cath-Ka (10 µM) for 4 h (for the detection of TLR2, TLR4, and MyD88) or for 0.5 h (for the detection of MAPKs) before the proteins of TLR2/4-MyD88-MAPKs signaling pathways were measured by WB. ( E ) Representative WB images (left) and quantitative analysis (right) of TLR2/4-MyD88-MAPKs signaling protein expression in peritoneal cells activated by Cath-Ka. Data are expressed as mean ± SD ( n = 3–4). * P < 0.05, ** P < 0.01, and *** P < 0.001 are thought statistically significant to the control group, while # P < 0.05, ## P < 0.01, and ### P < 0.001 are deemed to have significant differences between the corresponding Cath-Ka treatment groups; ns means not significant

Article Snippet: The inhibitors against TLR2 (C29, #HY-100461), TLR3 (CU-CPT 4a, #HY-108473), TLR4 (TAK-242, #HY-11109), TLR5 (TH1020, #HY-116961), TLR7/9 (Hydroxychloroquine, #HY-W031727), TLR7/8 (Enpatoran, #HY-134581), ERK (U0126, # HY-12031 A), p38 (SB203580H, # Y-10256), and JNK (SP600125, #HY-12041) were purchased from MedChem Express (Shanghai, China).

Techniques: Protein-Protein interactions, In Vitro, Expressing, Cell Culture, Control

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Cathelicidin-Ka, the first frog-derived TLR2 and TLR4 agonist, induces macrophage activation and promotes inflammation

doi: 10.1007/s00018-025-06068-y

Figure Lengend Snippet: Identification of the structure and function relation of Cath-Ka. ( A ) The interaction mode of Cath-Ka and TLR4 predicted by AlphaFold2 server and visualized by Pymol. ( B ) Hydrophobic interactions and hydrogen bond (green dashed line) of Cath-Ka-TLR4 complex around four residues at the N-terminus of Cath-Ka. ( C ) Comparison of their primary sequences and physicochemical parameters. The two cysteine of Cath-Ka-1 does not form the disulfide bond. ( D ) Secondary structure of Cath-Ka and its analogues in SDS solution. ( E - G ) Effect comparison of 10 µM Cath-Ka and its analogues on macrophage morphology ( E ), proliferation ( F ) and TNF-α production ( G ). Scale bar = 50 μm in Figure G. Data are expressed as mean ± SD ( n = 3). * : P < 0.05, ** : P < 0.01, and *** : P < 0.001 vs. Control; # : P < 0.05, ## : P < 0.01, and ### : P < 0.001 vs. Cath-Ka treatment; ns: Not significant

Article Snippet: The inhibitors against TLR2 (C29, #HY-100461), TLR3 (CU-CPT 4a, #HY-108473), TLR4 (TAK-242, #HY-11109), TLR5 (TH1020, #HY-116961), TLR7/9 (Hydroxychloroquine, #HY-W031727), TLR7/8 (Enpatoran, #HY-134581), ERK (U0126, # HY-12031 A), p38 (SB203580H, # Y-10256), and JNK (SP600125, #HY-12041) were purchased from MedChem Express (Shanghai, China).

Techniques: Comparison, Analogues, Control