lps-eb ultrapure  (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: 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: 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: Journal of Human Immunity

Article Title: Identification of a novel TLR7 gain-of-function variant that underlies systemic lupus erythematosus

doi: 10.70962/jhi.20250194

Figure Lengend Snippet: Evaluation of the L840R TLR7 variant function. Luciferase assay using HEK293T cells transfected with the pGL4.32 luciferase reporter construct and an expression vector for Renilla luciferase together with empty vector (Mock), or vectors expressing wild type (WT), LOF V795F TLR7 (negative control), GOF F507S and GOF L528I TLR7 (positive controls), or the L840R TLR7 variant stimulated or not by R848 TLR7 ligand. The results of four independent experiments are represented.

Article Snippet: After 24 h, the transfected cells were stimulated or not with 50 ng/mL R848 (Resquimod) for activation via TLR7/8 (Invivogen) for 24 h. Relative luciferase activity was then determined by normalizing the values against the firefly: Renilla luciferase signal ratio.

Techniques: Variant Assay, Luciferase, Transfection, Construct, Expressing, Plasmid Preparation, Negative Control

Journal: JID Innovations

Article Title: Dissecting the role of imiquimod and isostearic acid in Aldara-induced psoriasis mouse model

doi: 10.1016/j.xjidi.2026.100454

Figure Lengend Snippet: ISA does not cause psoriatic phenotype in C57BL/6 J mice. Dorsal skin of C57BL/6J mice were treated with Vaseline (control), 25% ISA, 62.5 mg of Aldara, 7.4% OA, or 62.5 mg of generic IMQ. ( a ) Representative images of mice at day 1, day 3, and day 6. ( b ) Dorsal skin was scored for erythema and scaling on a scale from 0 to 4 daily. Each point is representative of the mean score ± SEM (2-way repeated-measures ANOVA with Tukey’s posthoc test, n = 4). ∗ P < .05. ( c, d ) Histological sections of the dorsal skin were stained with H&E for acanthosis measurement (n = 4). Bar = 50 mm. ( d ) Average thickness of 3 separate measurements was taken from 3 different locations in the epidermis of each skin section (1-way ANOVA, Tukey’s multiple comparisons test, n = 4–5). ∗∗∗ P < .001. ctrl, control; IMQ, imiquimod; ISA, isostearic acid; ns, not significant; OA, oleic acid.

Article Snippet: IMQ was purchased from Invivogen.

Techniques: Control, Staining

Journal: JID Innovations

Article Title: Dissecting the role of imiquimod and isostearic acid in Aldara-induced psoriasis mouse model

doi: 10.1016/j.xjidi.2026.100454

Figure Lengend Snippet: ISA treatment does not cause systemic changes in C57BL/6 J mice. ( a ) Daily body weight changes of control, ISA-, Aldara-, OA-, and generic IMQ–treated mice represented as mean ± SEM (2-way repeated-measures ANOVA with Tukey’s posthoc test, n = 4). ∗ P < .05 and ∗∗∗ P < .001. ( b ) Splenocytes were assessed through flow cytometry for the presence of CD4 + , CD8 + , and CD11c + cells (1-way ANOVA, Tukey’s multiple comparisons test, n = 3–5). ∗∗ P < .01 and ∗∗∗ P < .001. ctrl, control; IMQ, imiquimod; ISA, isostearic acid; ns, not significant; OA, oleic acid.

Article Snippet: IMQ was purchased from Invivogen.

Techniques: Control, Flow Cytometry

Journal: JID Innovations

Article Title: Dissecting the role of imiquimod and isostearic acid in Aldara-induced psoriasis mouse model

doi: 10.1016/j.xjidi.2026.100454

Figure Lengend Snippet: ISA stimulation does not induce psoriasis signature inflammatory cytokines or pDC migration to the dermis. ( a ) Expression of IL-17A, IL-17F , and IL-1 β in the dorsal skin sections of control, ISA-, and Aldara-treated mice relative to the housekeeping gene L32 . Each data point is the mean of technical duplicates per mouse and represented as means ± SEM (1-way ANOVA, Dunnett’s multiple comparison test, n = 3–4). ∗ P < .05 and ∗∗ P < .01. ( b, c ) Immunofluorescence images of the dorsal skin of control, ISA-, and IMQ-treated mice stained with anti-PDCA1 and DAPI. Images were analyzed for cells in the dermis that were PDCA1 + DAPI + . Bar = 300 mm. ( c ) Cell counts were taken manually by 2 separate observers. Each point is representative of the mean cell count in the dermis per mouse. Data are represented as mean ± SEM (1-way ANOVA, Dunnett’s multiple comparisons test, n = 4–5). ∗∗ P < .01. ctrl, control; IMQ, imiquimod; ISA, isostearic acid; ns, not significant; pDC, plasmacytoid dendritic cell.

Article Snippet: IMQ was purchased from Invivogen.

Techniques: Migration, Expressing, Control, Comparison, Immunofluorescence, Staining, Cell Characterization

Journal: JID Innovations

Article Title: Dissecting the role of imiquimod and isostearic acid in Aldara-induced psoriasis mouse model

doi: 10.1016/j.xjidi.2026.100454

Figure Lengend Snippet: Treatment with isostearic acid does not induce activation or gene expression in pDCs in vitro. ( a ) CAL-1 cells were treated with ISA or IMQ for the hours indicated. Gene expressions of TNFα, IL-1β, and IL-6 were determined by quantitative real-time PCR using the ΔΔCt method. All values were normalized to the housekeeping gene L32 . Data are shown as technical duplicates from at least 3 separate experiments and represented as mean ± SEM (2-way ANOVA with Dunnett’s posthoc test). ∗∗∗ P < .001. ( b ) Cytoplasmic and nuclear extracts were prepared and analyzed by western blotting with antibodies for p65, p50, and c-Rel. β -tubulin and HDAC2 was used as loading controls, and hnRNPA1 and vinculin were used as purity controls for the cytoplasmic and nuclear fractions, respectively. IMQ, imiquimod; ISA, isostearic acid; min, minute; ns, not significant; pDC, plasmacytoid dendritic cell.

Article Snippet: IMQ was purchased from Invivogen.

Techniques: Activation Assay, Gene Expression, In Vitro, Real-time Polymerase Chain Reaction, Western Blot

Journal: Redox Biology

Article Title: Modification of the dermal matrix by senescence associated lipids and its functional consequence

doi: 10.1016/j.redox.2026.104069

Figure Lengend Snippet: Free reactive oxygen species affect basal and LPS induced gene expression on lipid modified matrices. A Bar charts represent relative gene expression of TNFA and IL1A normalized to B2M showing mean ± SD, n = 4. THP-1 cells were activated with 15 nM PMA, cultured on modified matrices for 24 h, and then stimulated for 2 h with human TNFα (100 ng/ml), Flagellin (0.25 μg/ml, TLR5), FSL-1 (0.5 μg/ml, TLR2, TLR6), and Poly(I:C) (20 μg/ml TLR3), respectively. B, C Bar charts show relative expression levels of macrophages treated with 5 mM N-Acetylcysteine (NAC) immediately after seeding and cultured for 24 h, followed by stimulation with 500 ng/ml LPS for 2 h, mean ± SD, n = 4. Asterisks indicate statistical significance (* p < 0.05; ** p < 0.01; *** p < 0.005; **** p < 0.001), and exact p-values are shown for borderline significance determined by one-way ANOVA.

Article Snippet: In detail, human TNFα (InvitrogenThermo Fisher, USA) was added to a final concentration of 100 ng/ml, 0.25 μg/ml of Flagellin (TLR5, InvivoGen, France), 0.5 μg/ml of FSL-1 (TLR2, TLR6, InvivoGen) and 20 μg/ml of Poly(I:C) (TLR3, InvivoGen) [ , ].

Techniques: Gene Expression, Modification, Cell Culture, Expressing