tlr ligands pam2csk4 pam2  (InvivoGen)

Journal: Journal of Translational Autoimmunity

Article Title: Type I interferon-dependent and -independent signaling underlie autoantibody production in a murine lupus model

doi: 10.1016/j.jtauto.2026.100351

Figure Lengend Snippet: Induction of SLE autoantibodies occurs in the presence of LPS or Poly I:C, but not other TLR ligands. Wild type (WT) C57BL/6 mice received five immunizations with β2GPI in combination with one of the following TLR ligands: TLR2 : PAM2, FSL-1 (lipoprotein); TLR3 : Poly I:C (dsRNA); TLR4 : LPS; TLR7: IMQ (imiquimod); or TLR9 : CpG-ODN (ssDNA). Serum autoantibodies were detected by ELISA. Bars indicate the mean value +/standard error (S.E.) per group (n = 5 mice/group), and dots indicate values for individual mice. Autoantibody levels for control mice immunized with PBS and TLR ligand were OD 405 < 0.1. The OD 405 + 3 S.D. for unimmunized mice was ≤0.1 for all autoantibodies assayed. (A) Anti-β2GPI and anti-CL antibodies, and (B) hallmark SLE autoantibodies (anti-DNA, anti-Ro/SS-A, anti-La/SS-B, and anti-Sm) were analyzed statistically in separate groupings. For each of the two autoantibody categories, each different TLR ligand (immunized with β2GPI) was compared to the other TLR ligands by 2-way ANOVA, followed by a Tukey's multiple comparison test. ns = not significant. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, and ∗∗∗∗P < 0.0001.

Article Snippet: TLR ligands Pam2CSK4 (PAM2), FSL-1, Pam3CSK4 (PAM3), Poly I:C (HMW), imiquimod-R837 (IMQ), and CpG-ODN-1555 + 1466 (CpG-ODN) were from InvivoGen (San Diego, CA).

Techniques: Enzyme-linked Immunosorbent Assay, Control, Comparison

Journal: bioRxiv

Article Title: A cilia-dependent inflammatory programme links bacterial detection to kidney disease

doi: 10.64898/2026.04.24.720658

Figure Lengend Snippet: (A) qPCR quantification of Ccl2 mRNA in Kif3a i-shRNA MDCK cells treated for 6 hours with TLR-1/2 (Pam3), TRL-2/6 (Pam2), TLR-4 (Lipopolysaccharide, LPS) or TLR-5 agonist (Flagellin, FLA). (B) qPCR quantification of Ccl2 mRNA in luciferase (blue), Kif3a (red) or Ift88 (green) i-shRNA MDCK cells treated for 6 hours by NOD1 (Tri-DAP) or NOD2 agonist (MDP). (C) qPCR quantification of Ccl2 mRNA in doxycycline (DOX) inducible luciferase (blue), Kif3a (red) or Ift88 (green) shRNA MDCK cells treated for 6 hours with 13µM ADPh. (D) qPCR quantification of Ccl2 mRNA in Kif3a i-shRNA MDCK cells treated for 4 hours with 500nM DF-006. (E-F) qPCR quantification of Alpk1 (E) and Tifa (F) mRNA in doxycycline (DOX) inducible luciferase (blue), Kif3a (red) or Ift88 (green) shRNA MDCK cells. (G) qPCR quantification of Ccl2 mRNA in Kif3a i-shRNA MDCK cells treated for 30min with 13µM ADPh in presence of digitonin, and stopped 5 hours and a half later. (H) Percentage of cells with TIFAsome formation in HeLa cells upon ADPh, UPEC heat-killed supernatant stimulation from WT (snUTI89 WT) or lacking Hlde (sn UTI89 ΔHlde), an essential enzyme to generate ADPh. (I-J) qPCR quantification of Ccl2 mRNA expression (expressed in percentage of the maximum value) depending on treatment duration (0min, 30min, 1h, 2h, 4h or 6h) after ADPh (I) and UPEC (J) stimulation of Kif3a i-shRNA MDCK cells. (K) qPCR quantification of Ccl2 mRNA in Kif3a i-shRNA MDCK cells treated for 6 hours with UPEC heat-killed supernatant lacking Hlde (sn UTI89 ΔHlde). (A-B, E-F) Ratio paired t test: *P<0.05, **P<0.01, ***P<0.001, ****P<0.00001. Each dot represents an experimental n. AU: arbitrary units. (D, G-H, K) Paired one-way RM ANOVA (log-normal) with Geisser Greenhouse correction followed by Tukey test: *P<0.05, **P<0.01, ***P<0.001, ****P<0.00001. Each dot represents an experimental n. AU: arbitrary units.

Article Snippet: Pam2 (tlrl-pm2s-1, InvivoGen) and Pam3 (tlrl-pms, InvivoGen) were dissolved in endotoxin-free water at 5 mg/ml and added to the cell culture medium at 5μg/mL final concentration during 1 hour of pre-treatment before stimulation.

Techniques: shRNA, Luciferase, Expressing