Journal: Microbiome

Article Title: An in vitro model demonstrating homeostatic interactions between reconstructed human gingiva and a saliva-derived multispecies biofilm.

doi: 10.1186/s40168-025-02033-w

Figure Lengend Snippet: Fig. 5 Immunohistochemical staining of RHG in the HMI model. Expression of elafin and hBD2 in RHG co-cultured with (+) or without (−) biofilms for 2 or 4 days. Representative staining pictures are shown, from four independent experiments, each with an intra-experimental replicate. Biofilm (−), biofilm-negative condition; biofilm (+), biofilm-positive condition; MB, McBain medium; AS, artificial saliva medium; FSM, filtered saliva plus mucin medium

Article Snippet: Tissue sections (5 μm) were cut and stained with hematoxylin and eosin (H&E) for histological analysis or used for immunohistochemistry with antibodies against Ki-67 (Dako, Agilent, USA), elafin (Hycult Biotech, The Netherlands), and human β-defensin 2 (hBD2) (OriGene Technologies, Rockville, USA), as previously described [41, 49].

Techniques: Immunohistochemical staining, Staining, Expressing, Cell Culture

Journal: Clinical proteomics

Article Title: Multi-omics analyses, cell experiments, and network pharmacology tools identified key proteins and candidate drugs for alopecia areata treatment.

doi: 10.1186/s12014-025-09544-6

Figure Lengend Snippet: Fig. 2 The figure contains the results of MR and Phe-MR analysis. The significance of causal association between 6 candidates of drug target and AA is evaluated by MR analysis on three independent cohorts (A). The results of the meta-analysis based on the statistics from MR are also shown in this forest plot (A). The dot plots illustrate the causal effects of identified drug targets, including DEFB1 (B), CYD5B2 (C), and HGFAC (D), on both dichotomous and quantitative phenotypes in the AstraZeneca database. The red dotted line represents the threshold for Bonferroni-corrected p-values

Article Snippet: The primary antibodies used were anti-β-actin (1:10000, Proteintech, 20536-1-AP), anti-GAPDH(1:10000, Proteintech, 60004-1-Ig), anti-DEFB1(1:1000, Proteintech, 14738-1-AP), anti-TGFβ1(1:5000, Proteintech, 81746- 2-RR), anti-SMAD2 (1:2000, Proteintech, 12570-1-AP), anti-p-SMAD2(1:1000, Proteintech, 80427-2-RR), and anti-MICA (1:1000, Proteintech, 66384-1-Ig).

Techniques:

Journal: Clinical proteomics

Article Title: Multi-omics analyses, cell experiments, and network pharmacology tools identified key proteins and candidate drugs for alopecia areata treatment.

doi: 10.1186/s12014-025-09544-6

Figure Lengend Snippet: Fig. 4 The figure shows the DEFB1 transfection efficiency in HaCaT cells at different timepoints after the transfection. The RNA (A) and protein (B) expres sion of DEFB1 both evaluated significantly after 48 h transfection with plasmid

Article Snippet: The primary antibodies used were anti-β-actin (1:10000, Proteintech, 20536-1-AP), anti-GAPDH(1:10000, Proteintech, 60004-1-Ig), anti-DEFB1(1:1000, Proteintech, 14738-1-AP), anti-TGFβ1(1:5000, Proteintech, 81746- 2-RR), anti-SMAD2 (1:2000, Proteintech, 12570-1-AP), anti-p-SMAD2(1:1000, Proteintech, 80427-2-RR), and anti-MICA (1:1000, Proteintech, 66384-1-Ig).

Techniques: Transfection, Plasmid Preparation

Journal: Clinical proteomics

Article Title: Multi-omics analyses, cell experiments, and network pharmacology tools identified key proteins and candidate drugs for alopecia areata treatment.

doi: 10.1186/s12014-025-09544-6

Figure Lengend Snippet: Fig. 5 The alteration of MICA expression and TGFβ/SMAD pathway after the DEFB1 overexpression. The RNA (A) and protein expression (B) of MICA both increased in HaCaT cells after the DEFB1 overexpression. The TGFβ/SMAD pathway was enriched in the AA samples of GSE212447 (C and D). The same pathway activation was also detected after the DEFB1 overexpression (E)

Article Snippet: The primary antibodies used were anti-β-actin (1:10000, Proteintech, 20536-1-AP), anti-GAPDH(1:10000, Proteintech, 60004-1-Ig), anti-DEFB1(1:1000, Proteintech, 14738-1-AP), anti-TGFβ1(1:5000, Proteintech, 81746- 2-RR), anti-SMAD2 (1:2000, Proteintech, 12570-1-AP), anti-p-SMAD2(1:1000, Proteintech, 80427-2-RR), and anti-MICA (1:1000, Proteintech, 66384-1-Ig).

Techniques: Expressing, Over Expression, Activation Assay

Journal: Clinical proteomics

Article Title: Multi-omics analyses, cell experiments, and network pharmacology tools identified key proteins and candidate drugs for alopecia areata treatment.

doi: 10.1186/s12014-025-09544-6

Figure Lengend Snippet: Fig. 8 The Cimigenol treatment inhibited the TGFβ/SMAD2 pathway activation and increased level of MICA induced by DEFB1 in vitro. (A) Cell viability of HaCaT cells in different concentrations of Cimigenol. (B) The Cimigenol treatment reversed the increase in the protein level of DEFB1, TGFβ1, p-SMAD2, and MICA after the transfection

Article Snippet: The primary antibodies used were anti-β-actin (1:10000, Proteintech, 20536-1-AP), anti-GAPDH(1:10000, Proteintech, 60004-1-Ig), anti-DEFB1(1:1000, Proteintech, 14738-1-AP), anti-TGFβ1(1:5000, Proteintech, 81746- 2-RR), anti-SMAD2 (1:2000, Proteintech, 12570-1-AP), anti-p-SMAD2(1:1000, Proteintech, 80427-2-RR), and anti-MICA (1:1000, Proteintech, 66384-1-Ig).

Techniques: Activation Assay, In Vitro, Transfection

Journal: Infection and Immunity

Article Title: Bacterial Periplasmic Oxidoreductases Control the Activity of Oxidized Human Antimicrobial β-Defensin 1

doi: 10.1128/IAI.00875-17

Figure Lengend Snippet: The antimicrobial activity of hBD1ox is specific for Gram-negative bacteria. HBD1ox and hBD1red were incubated with Gram-negative bacteria in an aerobic (A) or anaerobic (B) environment in the radial diffusion assay. Antimicrobial activity was measured by analyzing the diameter of the inhibition zone. A diameter of 2.5 mm (dotted line) is the diameter of the punched well. Data are presented as mean ± SEM from three independent experiments.

Article Snippet: Reduced hBD1 (hBD1red) was chemically synthesized by Emc Microcollections (Tuebingen, Germany).

Techniques: Activity Assay, Bacteria, Incubation, Diffusion-based Assay, Inhibition

Journal: Infection and Immunity

Article Title: Bacterial Periplasmic Oxidoreductases Control the Activity of Oxidized Human Antimicrobial β-Defensin 1

doi: 10.1128/IAI.00875-17

Figure Lengend Snippet: Antimicrobial activity against bacteria with mutations in the genes of interest. (A) Schematic overview of located membrane and periplasmic proteins in bacteria. (B to D) Two micrograms of hBD1ox and hBD1red was tested against E. coli strains with different protein knockouts in the outer membrane (B), cytosol (C), and periplasmic space (D) by radial diffusion assay. (D) HBD1ox shows a decreased antimicrobial activity against bacteria without DsbA or inner membrane proteins DsbB and TonB in comparison to the WT E. coli MC1000. The diameter of the inhibition zone was measured in the radial diffusion assay to determine the antimicrobial activity. A diameter of 2.5 mm (dotted line) is the diameter of the punched well. Results from experiments with wild-type E. coli MC1000 were pooled (n = 18) and used as the control for all the times when E. coli MC1000 mutants (n = 3) are assessed. All data are presented as mean ± SEM.

Article Snippet: Reduced hBD1 (hBD1red) was chemically synthesized by Emc Microcollections (Tuebingen, Germany).

Techniques: Activity Assay, Bacteria, Membrane, Diffusion-based Assay, Comparison, Inhibition, Control

Journal: Infection and Immunity

Article Title: Bacterial Periplasmic Oxidoreductases Control the Activity of Oxidized Human Antimicrobial β-Defensin 1

doi: 10.1128/IAI.00875-17

Figure Lengend Snippet: Bacterial oxidoreductases DsbA and DsbB are essential for the activity of hBD1ox. (A) Two micrograms of hBD1ox and hBD1red was tested against E. coli with protein double knockouts in the redox system and outer membrane. Bacteria without both oxidoreductases obtain resistance against hBD1ox (**, P = 0.0044; ****, P < 0.0001). (B) Membrane depolarization assay of E. coli MC1000 ΔdsbA ΔdsbB incubated with reduced and oxidized hBD1 for 1 h. HBD3ox (50 μg/ml) was used as positive control (+), and the negative control (−) was incubated without any peptide. HBD1ox is not able to significantly affect the bacterial membrane (**, P = 0.0067). (C) Four micrograms of hBD1ox was used in the plasmid induction experiment with either E. coli MC1000 ΔdsbA ΔdsbB, E. coli MC1000 ΔdsbA ΔdsbB harboring plasmids pQE60::dsbB and pBAD33::dsbA to generate the wild-type phenotype, or, as a control, E. coli MC1000 ΔdsbA ΔdsbB with empty plasmids pQE60 and pBAD33. Plasmid expression was during the 2.5 h of incubation, with addition of 0.4% l-arabinose and 2 mM IPTG every 30 min. HBD1ox shows antimicrobial activity against the mutant containing plasmids pQE60::dsbB and pBAD33::dsbA (**, P = 0.0036; ****, P < 0.0001). The diameter of the inhibition zone was measured in a radial diffusion assay to determine the antimicrobial activity. Results of representative radial diffusion assays are shown. A diameter of 2.5 mm (dotted line) is the diameter of the punched well. Data are presented as mean ± SEM from at least three independent biological replicates. Student's t test was used for comparison of normally distributed data.

Article Snippet: Reduced hBD1 (hBD1red) was chemically synthesized by Emc Microcollections (Tuebingen, Germany).

Techniques: Activity Assay, Membrane, Bacteria, Incubation, Positive Control, Negative Control, Plasmid Preparation, Control, Expressing, Mutagenesis, Inhibition, Diffusion-based Assay, Comparison

Journal: Bone Research

Article Title: A peptide containing the receptor binding site of insulin-like growth factor binding protein-2 enhances bone mass in ovariectomized rats

doi: 10.1038/s41413-018-0024-9

Figure Lengend Snippet: PEG-HBD1 peptide stimulates femoral aBMD in OVX rats. a Femoral aBMD values were obtained from different groups of rats using a DXA scanner at the beginning (W0), 4 weeks (W4), and 8 weeks (W8) of peptide treatment. Each data point represents mean ± SEM for each group animals at different time points. A significant difference was detected between sham ctrl rats and OVX ctrl rats at all three time points (indicated by the letter a). No difference was detected in all OVX rat treatment groups at baseline (W0) or after control peptide treatment at W4 and W8 time points (indicated by the letter b). Significant differences were detected between OVX rats treated with all three dose of PEG-HBD1 or PTH and OVX rats treated with a control peptide at W4 and W8 time points (indicated by the letter c, at least P < 0.05) except the mid dose of PEG-HBD1 at W8 time point. b Each bar graph represents mean ± SEM of each group of animals after 8 weeks of treatment. *** P < 0.001, ** P < 0.01, and * P < 0.05 indicated the significant difference between two treatments. c Each bar represents the percentage increase in femoral aBMD for that treatment compared to control (ctrl) peptide-treated OVX rats

Article Snippet: The synthetic peptide containing the heparin-binding domain 1 (HBD1) of human IGFBP-2 (CKHHLGLEEPKKLR) and a scrambled HBD1 peptide (CKPLRLSKEEHPLK) (control peptide) were synthesized by Genscript (Piscataway, NJ).

Techniques: Control

Journal: Bone Research

Article Title: A peptide containing the receptor binding site of insulin-like growth factor binding protein-2 enhances bone mass in ovariectomized rats

doi: 10.1038/s41413-018-0024-9

Figure Lengend Snippet: PEG-HBD1 peptide stimulates BV/TV of trabecular bone in the femur of OVX rats. a Representative images of femoral trabecular bone OVX rats treated with control (ctrl), high dose PEG-HBD1 peptide or PTH. b Each bar represents the mean ± SD of BV/TV of trabecular bone in femur from different group of animals. *** P < 0.001 and * P < 0.05 indicated the significant difference between two treatments. c Each bar represent the mean of BV/TV percentage change over control peptide (ctrl) treated rats by a different dose of peptide or PTH

Article Snippet: The synthetic peptide containing the heparin-binding domain 1 (HBD1) of human IGFBP-2 (CKHHLGLEEPKKLR) and a scrambled HBD1 peptide (CKPLRLSKEEHPLK) (control peptide) were synthesized by Genscript (Piscataway, NJ).

Techniques: Control

Journal: Bone Research

Article Title: A peptide containing the receptor binding site of insulin-like growth factor binding protein-2 enhances bone mass in ovariectomized rats

doi: 10.1038/s41413-018-0024-9

Figure Lengend Snippet: Histomorphometric analysis of the effect of the PEG-HBD1 peptide on bone volume/total volume (BV/TV) and trabecular thickness (TB.TH) of tibia in OVX rats. a , b Each bar graph represents the mean ± SD of BV/TV (a) or trabecular bone thickness ( b ) of tibia from the OVX rats with different treatments. ** P < 0.01 and * P < 0.05 indicates a significant difference between two treatments

Article Snippet: The synthetic peptide containing the heparin-binding domain 1 (HBD1) of human IGFBP-2 (CKHHLGLEEPKKLR) and a scrambled HBD1 peptide (CKPLRLSKEEHPLK) (control peptide) were synthesized by Genscript (Piscataway, NJ).

Techniques:

Journal: Bone Research

Article Title: A peptide containing the receptor binding site of insulin-like growth factor binding protein-2 enhances bone mass in ovariectomized rats

doi: 10.1038/s41413-018-0024-9

Figure Lengend Snippet: Histomorphometric analysis of the PEG-HBD1 peptide on mineral apposition rate (MAR) and bone formation rate (BFR) in tibia. a Representative images of demeclocycline and calcein staining, which showed the structural and dynamic change in the tibia from OVX rats after different treatments. b, c Each bar graph represents the mean ± SD of MAR ( b ) or BFR/bone surface (BS) ( c ) in the tibia from OVX rats with different treatments. * P < 0.05 indicates a significant difference between two treatments

Article Snippet: The synthetic peptide containing the heparin-binding domain 1 (HBD1) of human IGFBP-2 (CKHHLGLEEPKKLR) and a scrambled HBD1 peptide (CKPLRLSKEEHPLK) (control peptide) were synthesized by Genscript (Piscataway, NJ).

Techniques: Staining

Journal: Bone Research

Article Title: A peptide containing the receptor binding site of insulin-like growth factor binding protein-2 enhances bone mass in ovariectomized rats

doi: 10.1038/s41413-018-0024-9

Figure Lengend Snippet: PEG-HBD1 peptide increased osteoblasts number (N.Ob) and surface (Ob.S) but decreased osteoclasts surface (Oc.S) in tibia from OVX rats. Each bar graph represents the mean ± SD of N.Ob per bone perimeter (B.Pm) ( a ) or Ob.S/B.Pm ( b ) or Oc.S/B.Pm ( c ) in the tibia from OVX rats with different treatments. *** P < 0.001, ** P < 0.01, and * P < 0.05 indicated the significant difference between two treatments

Article Snippet: The synthetic peptide containing the heparin-binding domain 1 (HBD1) of human IGFBP-2 (CKHHLGLEEPKKLR) and a scrambled HBD1 peptide (CKPLRLSKEEHPLK) (control peptide) were synthesized by Genscript (Piscataway, NJ).

Techniques:

Journal: Bone Research

Article Title: A peptide containing the receptor binding site of insulin-like growth factor binding protein-2 enhances bone mass in ovariectomized rats

doi: 10.1038/s41413-018-0024-9

Figure Lengend Snippet: PEG- but not non PEG-HBD1 peptide had reverse dose response in stimulating osteocalcin expression in osteoblasts. a , b Cells were cultured as described in the materials and methods. Cell lysates were harvested and immunoblotted with an anti-osteocalcin antibody. The blots were reprobed with an anti-β-actin antibody as a loading control. The concentration of IGF-I was 100 ng·mL -1

Article Snippet: The synthetic peptide containing the heparin-binding domain 1 (HBD1) of human IGFBP-2 (CKHHLGLEEPKKLR) and a scrambled HBD1 peptide (CKPLRLSKEEHPLK) (control peptide) were synthesized by Genscript (Piscataway, NJ).

Techniques: Expressing, Cell Culture, Control, Concentration Assay