pgn sa tlr2  (InvivoGen)

Journal: bioRxiv

Article Title: Liver sinusoidal endothelial cells integrate metabolic and immune signals for MAPK-dependent BMP6 regulation and hepcidin induction

doi: 10.64898/2026.05.07.723498

Figure Lengend Snippet: (A) Bmp6 mRNA expression in primary LSECs treated with 5 ng/mL LPS or 2.5 μM heme for 6 h in the presence or absence of hepatocyte-conditioned medium. (B) Bmp6 mRNA expression in primary LSECs treated with various TLR ligands: Pam3CSK4 (TLR1/2), PGN (TLR2), Poly I:C (TLR3), LPS (TLR4), FLA-ST (TLR5), FSL1 (TLR2/6), R848 (TLR7/8), and ODN (TLR9) or vehicle control (NT, non-treated) for 6 h. (C-D) Bmp6 mRNA expression in primary LSECs treated with increasing concentrations of LPS for 6 h or with 5 ng/mL LPS for 2, 4, and 6 h. (E) Bmp6 mRNA expression in LSECs treated with 2.5 μM heme or protoporphyrin IX (PPIX) for 6 h. (F-G) Bmp6 mRNA expression in primary LSECs treated with increasing concentrations of heme for 6 h or treated with 2.5 μM heme for 2, 4, or 6 h. (H) Bmp6 mRNA expression in primary LSECs pre-treated with TAK242 (5 μM) or DMSO for 1 h, followed by 5 ng/mL LPS or 2.5 μM heme treatment for 6 h. (I) Transcription factor activity analysis by RNA-seq of LSECs treated with heme with respect to untreated controls (contrast-wise), in presence of hepatocyte-conditioned medium. (J) Bmp6 mRNA expression in primary LSECs pre-treated with CHX (5 μM) or DMSO for 1 h, followed by 5 ng/mL LPS treatment for 6 h. Cell culture experiments, except those in panel A, were always conducted in the presence of hepatocyte-conditioned medium. Gene expression levels were assessed by RT-qPCR, normalized to the housekeeping gene Rpl19 , and expressed as fold change relative to vehicle-treated controls. The dashed line (ut) represents the mRNA expression of LSECs treated with the conditions shown, in the absence of LPS or heme. Data are obtained from three or four independent experiments and displayed as mean ± SD. Statistical significance: *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001, one-way ANOVA or Student’s t-test. CM, conditioned medium; PPIX, protoporphyrin IX; CHX, cycloheximide.

Article Snippet: The TLR ligands Pam3CSK4 (TLR2:1) (#tlrl-pms), PGN-SA (TLR2) (#tlrl-pgns2), Poly I:C (TLR3) (#tlrl-picw), FLA-ST (TLR5) (#tlrl-stfla), FSL1 (TLR2:TLR6) (tlrl-fsl), R848 (TLR7:8) (#tlrl-r848-1), ODN (TLR9) (#tlrl-1826) and the MAPK inhibitor SP600125 (#tlrl-sp60) were purchased form Invivogen and diluted in PBS (TLR ligands) or DMSO (SP600125).

Techniques: Expressing, Control, Activity Assay, RNA Sequencing, Cell Culture, Gene Expression, Quantitative RT-PCR

Journal: Cell Reports

Article Title: Immunological Fingerprints of Controllers Developing Neutralizing HIV-1 Antibodies

doi: 10.1016/j.celrep.2019.12.087

Figure Lengend Snippet: IL-12 Signaling Is Required for Superior Tfh-Priming Properties of mDCs from Nt2 Controllers (A) Spearman correlation between frequencies of CXCR5 + PD-1 + pTfh from total CD4 + T cells and MFI of CD86 in mDCs from indicated controller Nts. Nt1 and Nt2 patients are highlighted in yellow and orange, respectively. R and p values for all Nts or all controllers are indicated in blue and black, respectively. (B) Box and whiskers plot showing the proportions of CXCR5 + PD-1 + Tfh-like cells induced from naive CD4 + T cells after 6 days of culture in the presence of autologous naive B cells and allogeneic mDCs from either Nt2 (orange, n = 5) or Nt1 (yellow, n = 5) Nts, NN (green, n = 9) controllers, and healthy donors (blue, n = 6). The error bars represent minimum to maximum values. Statistical significance was calculated using a two-tailed Mann-Whitney test. ∗∗ p < 0.01. (C) Box and whiskers plot showing the proportions of proliferating carboxyfluorescein succinimidyl ester low (CFSE low) (left panel) and viable (right panel) cells within CXCR5 + PD-1 + Tfh-like cells present in culture after incubation with mDCs from Nt2 (orange, n = 4 and 5) and Nt1 (yellow, n = 5 and 6) Nts, NNs (green, n = 9 and 10), and healthy donors (blue, n = 6). The error bars represent minimum to maximum values. Statistical significance was tested using a Mann-Whitney test. (D) Box and Whiskers plot showing proportions of proliferating CFSElow (left panel) and viable (right panel) cells within CXCR5 − PD-1 + non-Tfh cells present in culture after incubation with mDCs from Nt2 (orange, n = 4) and Nt1 (yellow, n = 5) Nts, NNs (green, n = 9), and healthy donors (blue, n = 5). Statistical significance was tested using a Mann-Whitney test. Error bars represent minimum to maximum values. (E) Proportions of IL-6 (left) and IL-12p70/p40 (right) producing cells in CD14 + Mo (top panels) and CD14 − CD11c Hi HLA-DR + mDCs (bottom panels) isolated from the blood of Nt2 (orange, n = 10), Nt1 (yellow, n = 6) Nts, and NN (green, n = 6) controllers cultured for 24 h in the presence of TLR8/7 or TLR2 agonists. The error bars represent minimum to maximum values. Values were corrected for basal levels present in media only (see Figure S5 ). The statistical significance between each condition among different patient cohorts was calculated using a Mann-Whitney test. ∗ p < 0.05. (F) Box and Whiskers plot showing proportions of CXCR5 + PD-1 + (left plot) and CXCR3 Hi PD-1 + (right plot) CD4 + T cells from healthy donors induced in the presence of mDCs from Nt2 controllers in the presence of DMSO (n = 7) or STAT4 (n = 7) inhibitors. ∗ p < 0.05, 2-tailed Wilcoxon matched-pairs signed rank test. Error bars represent minimum to maximum values. (G) Box plot and Whiskers plot showing fold change in the induction of Tfh-like cells defined as CXCR5 + PD-1 + (left panel) or Bcl-6 + PD-1 + cells (right panel) in the presence of mDCs isolated from healthy donors preincubated for 24 h in the presence of cytokines (IL-12 or IL-6 or TGF-β, n = 6 each) or TLR ligands (PGNSA or poly I:C, n = 6, or lipopolysaccharide [LPS], n = 5). Data were normalized to culture conditions with unstimulated mDCs. The error bars represent minimum to maximum values. ∗ p < 0.05, two-tailed Wilcoxon matched-pairs signed rank test.

Article Snippet: PGN-SA TLR2 Agonist , InvivoGen , tlrl-pgns2.

Techniques: Two Tailed Test, MANN-WHITNEY, Incubation, Isolation, Cell Culture

Journal: Cell Reports

Article Title: Immunological Fingerprints of Controllers Developing Neutralizing HIV-1 Antibodies

doi: 10.1016/j.celrep.2019.12.087

Figure Lengend Snippet:

Article Snippet: PGN-SA TLR2 Agonist , InvivoGen , tlrl-pgns2.

Techniques: Staining, Plasmid Preparation, Recombinant, Isolation, Sequencing, Sample Prep, Cell Isolation, Expressing, Software