Journal: PLoS Pathogens

Article Title: A human secretome library screen reveals a role for Peptidoglycan Recognition Protein 1 in Lyme borreliosis

doi: 10.1371/journal.ppat.1009030

Figure Lengend Snippet: (A) Schematic of yeast-display screen- A library of yeast cells, each displaying a single human protein encoded by a uniquely bar-coded plasmid was pooled and mixed with surface biotinylated Borrelia . The BASEHIT library was scaled to 96-well magnetic separation format for this screen. Magnetic separation using streptavidin microbeads, followed by next-generation sequencing was used to identify yeast displaying proteins that bind to the bacterial cells. Plasmid DNA was isolated and sequenced to identify the proteins (B) Host interactions with B . burgdorferi N40- spirochete from the cultures grown at 33°C or 37°C were surface biotinylated and used for yeast selections, as described above. The data shows average scores of four independent runs of the selection on the 33°C grown sample and one run on the 37°C grown sample. The scores were normalized for diverse microbes (i.e. Escherichia coli , Staphylococcus aureus , Bacillus subtilis , Shigella flexneri , and Bifidobacterium adolescentis ) as background correction for non-specific binding activity. (C) PGLYRP1 interaction with Borrelia species- Samples from 53 Borrelia isolates, grown at two different temperatures (when possible), were screened against the host protein library as described above for B . burgdorferi , as were 370 additional bacterial samples. The score for each gene is defined as the overall enrichment for that gene (relative to the unselected library) multiplied by the percentage of barcodes associated with the gene that enriched (defined as logFC >0). The calculated scores of Borrelia species were compared with those of the other bacteria. The bars represent mean ± SD and p-values were determined using a Mann-Whitney U-test.

Article Snippet: Low passage B . burgdorferi were cultured to a density of ~10 6 −10 7 cells/mL, washed two times with PBS and incubated with either recombinant human PGLYRP1 (with 8X-His tag), recombinant mouse PGLYRP1 (with 6X-His tag) (Sino Biological, #50115-M08H), recombinant human CD55 (with 8X-His tag), or murine CD55 (with 8X-His tag) at room temperature for 1 hour.

Techniques: Plasmid Preparation, Next-Generation Sequencing, Isolation, Selection, Binding Assay, Activity Assay, MANN-WHITNEY

Journal: PLoS Pathogens

Article Title: A human secretome library screen reveals a role for Peptidoglycan Recognition Protein 1 in Lyme borreliosis

doi: 10.1371/journal.ppat.1009030

Figure Lengend Snippet: (A) ELISA results show the interaction of human PGLYRP1 with lysate of B . burgdorferi . The lysate was immobilized on microtiter wells and probed with increasing concentrations of either recombinant human PGLYRP1-Fc protein (1–100 ng, red) or Fc control protein (gray). The values plotted represent the mean ± SEM of three replicates from a single experiment. p-value is displayed in the graph and determined using a student t-test. (B and C) Binding of human (B) and mouse (C) PGLYRP1 to B . burgdorferi . The culture was grown to a density of 10 6 CFU/mL and incubated with varying concentrations of recombinant PGLYRP1-His 8 (10 μg/mL in green, 40 μg/ml in red). B . burgdorferi bound to recombinant PGLYRP1-His 8 was measured using a secondary AF488-His 6 monoclonal antibody by flow cytometry. Overlay histograms show protein binding to B . burgdorferi identified by Alexa Fluor 488-His 6 monoclonal antibody. Binding of recombinant human/murine CD55-His 8 (40 μg/ml in blue) to B . burgdorferi was used as control. The background binding of AF488-His 6 antibody alone with B . burgdorferi is shown in gray shaded region. Results from one independent experiment are shown here. For (B) and (C), the Y-axis represents relative cell counts calculated as a percentage of the maximum events ( Borrelia ).

Article Snippet: Low passage B . burgdorferi were cultured to a density of ~10 6 −10 7 cells/mL, washed two times with PBS and incubated with either recombinant human PGLYRP1 (with 8X-His tag), recombinant mouse PGLYRP1 (with 6X-His tag) (Sino Biological, #50115-M08H), recombinant human CD55 (with 8X-His tag), or murine CD55 (with 8X-His tag) at room temperature for 1 hour.

Techniques: Enzyme-linked Immunosorbent Assay, Recombinant, Binding Assay, Incubation, Flow Cytometry, Protein Binding

Journal: PLoS Pathogens

Article Title: A human secretome library screen reveals a role for Peptidoglycan Recognition Protein 1 in Lyme borreliosis

doi: 10.1371/journal.ppat.1009030

Figure Lengend Snippet: (A) ELISA shows the binding of recombinant human PGLYRP1 at two concentrations, 100 and 500 ng/ml to peptidoglycan (PG) sacculi isolated from B . burgdorferi (Bb) as compared to a PBS negative control. The values plotted represent the mean ± SEM of two technical replicates from a single experiment. (B and C) Flow cytometry-based experiment showing binding of human (B) and mouse (C) PGLYRP1 to B . burgdorferi , after pre-incubating the protein in the absence (red) and presence (blue) of B . burgdorferi PG. Recombinant PGLYRP1-His 6 (1 μg/mL) was pre-incubated with Borrelia PG (10 μg/mL) and then added to live B . burgdorferi overlay histograms show PGLYRP1 binding to B . burgdorferi identified by Alexa Fluor 488, His 6 monoclonal antibody. Results from one independent experiment are shown here. For (B) and (C), the Y-axis represents relative cell counts calculated as a percentage of the maximum events ( Borrelia ).

Article Snippet: Low passage B . burgdorferi were cultured to a density of ~10 6 −10 7 cells/mL, washed two times with PBS and incubated with either recombinant human PGLYRP1 (with 8X-His tag), recombinant mouse PGLYRP1 (with 6X-His tag) (Sino Biological, #50115-M08H), recombinant human CD55 (with 8X-His tag), or murine CD55 (with 8X-His tag) at room temperature for 1 hour.

Techniques: Enzyme-linked Immunosorbent Assay, Binding Assay, Recombinant, Isolation, Negative Control, Flow Cytometry, Incubation

Journal: PLoS Pathogens

Article Title: A human secretome library screen reveals a role for Peptidoglycan Recognition Protein 1 in Lyme borreliosis

doi: 10.1371/journal.ppat.1009030

Figure Lengend Snippet: Wild-type BALB/c and PGLYRP1 -/- mice (at least n = 7 in each group) were infected with 1x10 6 spirochetes by subcutaneous injection. (A and B) Skin B . burgdorferi burden was assessed by ear punch biopsies at 14 d (A) and 25 d (B) post infection, by qPCR for Borrelia specific gene ( flaB ) normalized to mouse β-actin . (C and D) B . burgdorferi burden was assessed in hearts (C) and joints (D) at day 25, by qPCR as above. Results from two independent experiments are shown. Each data point represents the value of an individual animal. (E) The extent of splenomegaly was expressed as spleen weights in WT and PGLYRP1 -/- mice infected with B . burgdorferi at day 25. Results from one independent experiment are shown here. Each data point represents the value of an individual animal. The bars represent mean ± SEM and p-values are calculated by student t-test.

Article Snippet: Low passage B . burgdorferi were cultured to a density of ~10 6 −10 7 cells/mL, washed two times with PBS and incubated with either recombinant human PGLYRP1 (with 8X-His tag), recombinant mouse PGLYRP1 (with 6X-His tag) (Sino Biological, #50115-M08H), recombinant human CD55 (with 8X-His tag), or murine CD55 (with 8X-His tag) at room temperature for 1 hour.

Techniques: Infection, Injection

Journal: PLoS Pathogens

Article Title: A human secretome library screen reveals a role for Peptidoglycan Recognition Protein 1 in Lyme borreliosis

doi: 10.1371/journal.ppat.1009030

Figure Lengend Snippet: (A) Histopathology scores from tibiotarsi for individual mice in infected wild-type mice (WT Inf) and infected PGLYRP1 -/- mice (PGLYRP1 -/- Inf) at 25 days post infection. Tibiotarsi were scored by blinded examination for tenosynovitis on a scale of 0 (negative) to 3 (severe). (B) The severity of cardiac inflammation in the heart of infected WT and PGLYRP1 -/- mice 25 d post infection. Hearts were scored in a blinded fashion for carditis on a scale of 0 (negative) to 5 (severe). Similar to tenosynovitis scores, PGLYRP1 -/- infected mice demonstrated no significant difference in carditis scores compared to WT mice. Results from at least two independent experiments (at least n = 7 in each group) are pooled and shown here. The bars represent mean ± SEM and p-values were calculated by Mann-Whitney U-test.

Article Snippet: Low passage B . burgdorferi were cultured to a density of ~10 6 −10 7 cells/mL, washed two times with PBS and incubated with either recombinant human PGLYRP1 (with 8X-His tag), recombinant mouse PGLYRP1 (with 6X-His tag) (Sino Biological, #50115-M08H), recombinant human CD55 (with 8X-His tag), or murine CD55 (with 8X-His tag) at room temperature for 1 hour.

Techniques: Histopathology, Infection, MANN-WHITNEY

Journal: PLoS Pathogens

Article Title: A human secretome library screen reveals a role for Peptidoglycan Recognition Protein 1 in Lyme borreliosis

doi: 10.1371/journal.ppat.1009030

Figure Lengend Snippet: Antibody levels in uninfected wild type BALB/c (WT) and PGLYRP1 -/- mice were compared with those in the infected ones (at least n = 7 in each group). Representative results from one independent experiment are shown. (A) Whole cell lysate of B . burgdorferi was coated on microtiter plate and serum from either uninfected WT, infected WT, uninfected PGLYRP1 -/- or infected PGLYRP1 -/- mice was used at varying dilutions. The binding was measured by secondary goat anti-mouse IgG HRP-conjugated antibody. Statistically significant increase in binding was observed at IgG titers 1:200, 1:2000, 1:20000 in infected WT compared to infected knockout mice. (B-E) Levels of different IgG isotypes (IgG1, B; IgG2a, C; IgG2b, D; IgG3, E) were measured against B . burgdorferi lysate, using mouse serum at varying dilutions. The binding was measured by secondary goat anti-mouse IgG1, IgG2a, IgG2b or IgG3 HRP-conjugated antibody. Statistically significant increase was observed in IgG1, IgG2a, IgG3, and IgG2b at 1:200 and 1:2000 dilution in infected WT compared to infected PGLYRP1 -/- mice. Representative results from one independent experiment are shown. Each data point represents an individual animal in the corresponding group. The bars represent mean ± SEM and p values determined using student t test.

Article Snippet: Low passage B . burgdorferi were cultured to a density of ~10 6 −10 7 cells/mL, washed two times with PBS and incubated with either recombinant human PGLYRP1 (with 8X-His tag), recombinant mouse PGLYRP1 (with 6X-His tag) (Sino Biological, #50115-M08H), recombinant human CD55 (with 8X-His tag), or murine CD55 (with 8X-His tag) at room temperature for 1 hour.

Techniques: Infection, Binding Assay, Knock-Out

Journal: PLoS Pathogens

Article Title: A human secretome library screen reveals a role for Peptidoglycan Recognition Protein 1 in Lyme borreliosis

doi: 10.1371/journal.ppat.1009030

Figure Lengend Snippet: Serum cytokine profile was assessed in both wild-type and PGLYRP1 -/- mice 25 d post-infection ((at least n = 7 in each group) using a mouse cytokine/chemokine 31-plex (MD-31) array. An increase in pro-inflammatory cytokines IFN-γ (A) and related cytokines- CXCL-9 (B) and CXCL-10 (C) was observed in infected PGLYRP1 -/- mice as compared to parent BALB/c mice. Representative results from one independent experiment are shown. Each data point represents an individual animal in the corresponding group. The bars represent mean ± SEM and p-values determined using student t test.

Article Snippet: Low passage B . burgdorferi were cultured to a density of ~10 6 −10 7 cells/mL, washed two times with PBS and incubated with either recombinant human PGLYRP1 (with 8X-His tag), recombinant mouse PGLYRP1 (with 6X-His tag) (Sino Biological, #50115-M08H), recombinant human CD55 (with 8X-His tag), or murine CD55 (with 8X-His tag) at room temperature for 1 hour.

Techniques: Infection

Journal: PLoS Pathogens

Article Title: A human secretome library screen reveals a role for Peptidoglycan Recognition Protein 1 in Lyme borreliosis

doi: 10.1371/journal.ppat.1009030

Figure Lengend Snippet: B . burgdorferi is a diderm bacteria where the outer membrane surrounds the peptidoglycan layer and protects it from the external environment. The peptidoglycan meshwork, in turn, surrounds the cytoplasm. Our results suggest PGLYRP1 binds to Borrelia peptidoglycan.

Article Snippet: Low passage B . burgdorferi were cultured to a density of ~10 6 −10 7 cells/mL, washed two times with PBS and incubated with either recombinant human PGLYRP1 (with 8X-His tag), recombinant mouse PGLYRP1 (with 6X-His tag) (Sino Biological, #50115-M08H), recombinant human CD55 (with 8X-His tag), or murine CD55 (with 8X-His tag) at room temperature for 1 hour.

Techniques:

Journal: Aging (Albany NY)

Article Title: PGC-1α activator ZLN005 promotes maturation of cardiomyocytes derived from human embryonic stem cells

doi: 10.18632/aging.103088

Figure Lengend Snippet: The expression of PGC-1α was upregulated during cardiomyocyte differentiation; ZLN005 increased PGC-1α mRNA and protein level in hESC-CMs. ( A ) The relative mRNA and ( B ) protein expression of PGC-α during cardiomyocyte differentiation (mRNA, n=7; protein, n=5). ( C ) Schematic representation of the experimental schedule including hESC culture, cardiomyocyte differentiation, culture and treatment. ( D ) Effect of ZLN005 on mRNA levels (n=6). ( E ) Effect of ZLN005 on PGC-α protein expression (n=12).

Article Snippet: The primary antibodies were as follows: anti-PGC-1α antibody (1:500, Boster biological), Connexin 43 antibody (1:400, Boster biological), and anti-GAPDH antibody (1:5000, Abcam).

Techniques: Expressing

Journal: Aging (Albany NY)

Article Title: PGC-1α activator ZLN005 promotes maturation of cardiomyocytes derived from human embryonic stem cells

doi: 10.18632/aging.103088

Figure Lengend Snippet: The used primers in the study.

Article Snippet: The primary antibodies were as follows: anti-PGC-1α antibody (1:500, Boster biological), Connexin 43 antibody (1:400, Boster biological), and anti-GAPDH antibody (1:5000, Abcam).

Techniques: Sequencing

Journal: Nature Communications

Article Title: PGLYRP1-mediated intracellular peptidoglycan detection promotes intestinal mucosal protection

doi: 10.1038/s41467-025-57126-9

Figure Lengend Snippet: a Peptidoglycan fragment library: GMTriP-K (1) and MDP (2); synthetic peptidoglycan fragments (1a-b, 2a-h and 3a-d) were all prepared with amine linkage points for attachment to the array surface via NHS-chemistry. b General workflow for the printing, incubation, and analysis of the PGN small fragment microarray (Created in BioRender. Putnik, R. (2025) https://BioRender.com/x50o125 ). c Microarray binding of PGLYRPs. PGLYRP1 binds to the disaccharide components of the array, whereas PGLYRP3 and PGLYRP4 show no association with the compounds on the array. d Determination of Apparent Dissociation Constant for PGLYRP1 to GMTriP-K (3 independent experiments were carried out with each experiment done in technical replicates). e Alphafold protein structure prediction of human PGLYRP1. f – i Docking prediction of GMTriP-K to human PGLYRP1. j Docking-prediction of PGLYRP3 and GMTriP-K demonstrating shallow GlcNAc preventing interaction. For synthetic procedures and compound characterization (NMRs, HRMS), please see the . For ( c ), each condition was screened in at least biological triplicate and technical replicated (see for raw image files and other biologically replicate binding data). Data are presented as mean ± SEM. All experiments were repeated twice and yielded consistent results.

Article Snippet: After washing the slides with PBS, the sections were incubated overnight in a primary antibody against PGLYRP1 (R&D Systems, #AF2696; 1:500 dilution), CD68 (Cell Signaling Technology, #76437; 1:1000 dilution), SEC61β (Cell Signaling Technology, # 14648 1:1000) and F4/80 (Cell Signaling Technology, #70076; 1:1000) in 4 degrees C in a humidified shielded chamber.

Techniques: Incubation, Microarray, Binding Assay

Journal: Nature Communications

Article Title: PGLYRP1-mediated intracellular peptidoglycan detection promotes intestinal mucosal protection

doi: 10.1038/s41467-025-57126-9

Figure Lengend Snippet: a RNA-seq and volcano plot analysis of gene expression in BMDMs from wild type and Pglyrp1 -/- mice after 18 h of stimulation with 25 µM GMTriP-K. b Volcano plot analysis of gene expression in BMDMs from wild type and NOD2 -/- mice after 18 h of stimulation with 25 µM GMTriP-K. c Volcano plot analysis of gene expression in BMDMs from wild type mice after 18 h of stimulation with 25 µM GMTriP-K or 25 µM MDP. d Hierarchal cluster analysis of genes induced by either stimulus in wild type or Pglyrp1 -/- BMDMs. e Hierarchical cluster analysis of genes induced by GMTriP-K in BMDMs from wild type, Pglyrp1 -/-, Nod2-/- , or Arhgef2 -/- mice. f qPCR analysis of gene expression in BMDMs from indicated mouse strains stimulated with 25 µM GMTriP-K or 25 µM GMDiP stimulation on BMDMs from wild type and Pylyrp1-/- mice for 18 h ( n = 3). g , h Gene set enrichment analysis of the differentially expressed genes induced by indicated stimulus in wild type BMDM. g Top 25 enriched signaling pathways for the differentially expressed gene sets. h Gene set enrichment analysis summarized in mountain plots representing significantly enriched (left) or depleted (right) of genes for the indicated gene sets and collections. Data are presented as mean ± SEM with indicated P values analyzed by one-way ANOVA. All experiments were repeated twice and yielded consistent results.

Article Snippet: After washing the slides with PBS, the sections were incubated overnight in a primary antibody against PGLYRP1 (R&D Systems, #AF2696; 1:500 dilution), CD68 (Cell Signaling Technology, #76437; 1:1000 dilution), SEC61β (Cell Signaling Technology, # 14648 1:1000) and F4/80 (Cell Signaling Technology, #70076; 1:1000) in 4 degrees C in a humidified shielded chamber.

Techniques: RNA Sequencing, Gene Expression, Protein-Protein interactions

Journal: Nature Communications

Article Title: PGLYRP1-mediated intracellular peptidoglycan detection promotes intestinal mucosal protection

doi: 10.1038/s41467-025-57126-9

Figure Lengend Snippet: a Hierarchal clustering of MDP induced genes in BMDMs from wild-type, Pglyrp1 -/- , Nod2 -/- , or Arhgef2 -/- mice. b , c BMDMs from wild type, Pglyrp1 -/ , Nod2 -/- , or Arhgef2 -/- mice were stimulated with 25 μM GMTriP-K or 25 μM MDP and analyzed via western blotting with the indicated antibodies. d BMDMs from wild type or Ifnar1 -/- mice were stimulated with 25 μM GMTriP-K or 25 μM MDP for 18 h and analyzed for STAT1 pathway activation with phospho-specific antibodies as indicated. e Wild type or GEF-H1-deficient BMDMs were treated with 100 ng/mL of IFNγ or IFNβ for 18 h and analyzed via western blotting with the indicated antibodies. f BMDMs from wild type, Pglyrp1 -/ , and Nod2 -/- mice were stimulated with 25 μM GMTriP-K or 25 μM MDP, lysed, and analyzed via western blotting with the indicated antibodies. g qPCR analysis of gene expression in BMDMs from indicated mouse strains stimulated with 25 µM GMTriP-K or 25 µM GMDiP for 18 h. Data are presented as mean ± SEM with indicated P values analyzed by one-way ANOVA. All experiments were repeated twice and yielded consistent results.

Article Snippet: After washing the slides with PBS, the sections were incubated overnight in a primary antibody against PGLYRP1 (R&D Systems, #AF2696; 1:500 dilution), CD68 (Cell Signaling Technology, #76437; 1:1000 dilution), SEC61β (Cell Signaling Technology, # 14648 1:1000) and F4/80 (Cell Signaling Technology, #70076; 1:1000) in 4 degrees C in a humidified shielded chamber.

Techniques: Western Blot, Activation Assay, Gene Expression

Journal: Nature Communications

Article Title: PGLYRP1-mediated intracellular peptidoglycan detection promotes intestinal mucosal protection

doi: 10.1038/s41467-025-57126-9

Figure Lengend Snippet: a – k Representative confocal microscopy images of HEK 293T cells expressing untagged (UT) or Flag-tagged PGLYRP1, orange fluorescent protein (OFP)-tagged SEC61β and GFP-tagged NOD2. Cells were transfected with PGLYRP1-UT ( a – d ) or PGLYRP1-Flag ( e – k ) and SEC61β-OFP and then probed with PGLYRP1 and GM130 antibodies. PGLYRP1, SEC61β-OFP, and GM130 were detected in Alexa Fluor 488 (green pseudo-color), mOrange (red), and Alexa Fluor 647 (blue) channels, respectively. l – y Cells were transfected with PGLYRP1-UT and NOD2-GFP and stained for PGLYRP1 and GM130. NOD2-GFP, PGLYRP1, and GM130 were detected in EGFP (green pseudo-color), Rhodamine Red-X (red), and Alexa Fluor 647 (blue) channels, respectively. All channels were scanned sequentially, and images are pseudo-colored independent of channel wavelengths with nuclear counterstain in greyscale (Scale bars indicate 2 μm in ( a – d, l – r ) and 5 μm in ( e – k , s – y )). Typically, 10-12 cells were analyzed for each condition. Data are presented as mean ± SEM Pearson’s coefficients were calculated in Imaris 10.1.1 image analysis software. All experiments were repeated twice and yielded consistent results.

Article Snippet: After washing the slides with PBS, the sections were incubated overnight in a primary antibody against PGLYRP1 (R&D Systems, #AF2696; 1:500 dilution), CD68 (Cell Signaling Technology, #76437; 1:1000 dilution), SEC61β (Cell Signaling Technology, # 14648 1:1000) and F4/80 (Cell Signaling Technology, #70076; 1:1000) in 4 degrees C in a humidified shielded chamber.

Techniques: Confocal Microscopy, Expressing, Transfection, Staining, Software

Journal: Nature Communications

Article Title: PGLYRP1-mediated intracellular peptidoglycan detection promotes intestinal mucosal protection

doi: 10.1038/s41467-025-57126-9

Figure Lengend Snippet: a Western blot analysis of protein expression in BMDMs from wildtype, Arhgef2-/- , Pglyrp1-/- , and Nod2-/- mice. b Time course of protein interactions with GEF-H1 after GMTriP-K stimulation of BMDMs. c Analysis of protein interactions with PGLYRP1 or d NOD2 after GMTriP-K stimulation of BMDMs. e Input assessment of protein expression for the IPs in ( c , d ). f Assessment of protein interactions with GEF-H1 after stimulation with indicated immune stimuli. Representative experiments of at least three repeats are shown.

Article Snippet: After washing the slides with PBS, the sections were incubated overnight in a primary antibody against PGLYRP1 (R&D Systems, #AF2696; 1:500 dilution), CD68 (Cell Signaling Technology, #76437; 1:1000 dilution), SEC61β (Cell Signaling Technology, # 14648 1:1000) and F4/80 (Cell Signaling Technology, #70076; 1:1000) in 4 degrees C in a humidified shielded chamber.

Techniques: Western Blot, Expressing

Journal: Nature Communications

Article Title: PGLYRP1-mediated intracellular peptidoglycan detection promotes intestinal mucosal protection

doi: 10.1038/s41467-025-57126-9

Figure Lengend Snippet: a Weight development of wild type and Pglyrp1 -/- mice during TNBS-induced colitis ( n = 4 per group). Immunofluorescence staining of PGLYRP1 and macrophages (F4/80) in colon sections of wild type and Pglyrp1 -/- mice before ( b ), after TNBS induced colitis ( c , d ) TNBS colitis induction after 3 days of i.p. pretreatment with 100 μg of GMTriP-K in 50 μL PBS at day 9. e Quantitation of F4/80 + macrophages and percentage of PGLYRP1 expressing F4/80 + macrophages during TNBS colitis. Macrophages were counted in 4 independent regions per section and quantitated using Imaris 10.1.1 image analysis software. f Staining for PGLYRP1 and SEC61β in the colon of wild type mice with TNBS colitis. g qPCR analysis of gene expression in mesenteric lymph nodes of wild type and Pglyrp1 -/- mice on day 9 of the TNBS and GMTPriP-K treatment protocol ( n = 3–5). Scale bars represent 20 μm in ( b , c , d) and 2 μm in ( g ). Data are presented as mean ± SEM with indicated P values analyzed by one-way ANOVA. All experiments were repeated twice and yielded consistent results.

Article Snippet: After washing the slides with PBS, the sections were incubated overnight in a primary antibody against PGLYRP1 (R&D Systems, #AF2696; 1:500 dilution), CD68 (Cell Signaling Technology, #76437; 1:1000 dilution), SEC61β (Cell Signaling Technology, # 14648 1:1000) and F4/80 (Cell Signaling Technology, #70076; 1:1000) in 4 degrees C in a humidified shielded chamber.

Techniques: Immunofluorescence, Staining, Quantitation Assay, Expressing, Software, Gene Expression

Journal: Nature Communications

Article Title: PGLYRP1-mediated intracellular peptidoglycan detection promotes intestinal mucosal protection

doi: 10.1038/s41467-025-57126-9

Figure Lengend Snippet: a Immunofluorescence staining of PGLYRP1 and macrophages (CD68) in tissue sections of healthy human colon and active ulcerative colitis (arrow indicates immune cells that crossed the epithelial barrier, cr, Crypt.). b Quantitation of CD68 + macrophages and percentage of PGLYRP1 expressing CD68 + macrophages in the normal colon and ulcerative colitis lamina propria. Macrophages were counted in 4 independent regions per sample and quantitated using Imaris 10.1.1 image analysis software. c Hierarchal cluster analysis of genes that were significantly regulated in active, inactive ulcerative colitis and healthy control colons. d Volcano plot analysis of gene expression comparing gene expression in healthy control and active ulcerative colitis. e qPCR analysis of expression of indicated genes in healthy control ( n = 6), inactive ( n = 8), and active ulcerative colitis ( n = 6) samples. Scale bars represent 25 μm. Data are presented as mean ± SEM with indicated P values analyzed by one-way ANOVA. Student’s test was used to compare control and UC samples. All experiments were repeated at least twice and yielded consistent results.

Article Snippet: After washing the slides with PBS, the sections were incubated overnight in a primary antibody against PGLYRP1 (R&D Systems, #AF2696; 1:500 dilution), CD68 (Cell Signaling Technology, #76437; 1:1000 dilution), SEC61β (Cell Signaling Technology, # 14648 1:1000) and F4/80 (Cell Signaling Technology, #70076; 1:1000) in 4 degrees C in a humidified shielded chamber.

Techniques: Immunofluorescence, Staining, Quantitation Assay, Expressing, Software, Control, Gene Expression

Journal: Nature Communications

Article Title: PGLYRP1-mediated intracellular peptidoglycan detection promotes intestinal mucosal protection

doi: 10.1038/s41467-025-57126-9

Figure Lengend Snippet: PGLYRP1 is required for detection of GMTriP-K. PGLYRP1 localizes to the endoplasmic reticulum and Golgi and interacts with NOD2 and GEF-H1 to induce the expression of immune regulators that regulate intestinal inflammation (Created in BioRender. Reinecker, H. (2025) https://BioRender.com/i55t156 ).

Article Snippet: After washing the slides with PBS, the sections were incubated overnight in a primary antibody against PGLYRP1 (R&D Systems, #AF2696; 1:500 dilution), CD68 (Cell Signaling Technology, #76437; 1:1000 dilution), SEC61β (Cell Signaling Technology, # 14648 1:1000) and F4/80 (Cell Signaling Technology, #70076; 1:1000) in 4 degrees C in a humidified shielded chamber.

Techniques: Expressing