Journal: Frontiers in Immunology

Article Title: Clinical and immunological characterization of a Netherton syndrome infant with a large SPINK gene cluster deletion and a c.1258A>G polymorphism in SPINK5

doi: 10.3389/fimmu.2025.1658444

Figure Lengend Snippet: Genetic and protein expression analysis. (A) Whole-exome sequencing identified a heterozygous deletion spanning the SPINK gene cluster, including SPINK5 . (B) Sanger sequencing of the patient and his mother revealed a polymorphism in SPINK5 ( NM_006846 ; exon 14, c.1258A>G, p.K420E). (C) LEKTI and KLK5 expression in peripheral blood mononuclear cells (PBMCs) from the patient compared to healthy controls (HC, n=3). (D) KLK5 level in the patient’s plasma (seven time points from initial hospitalization) compared with that in healthy controls (HC, n = 8).

Article Snippet: Membranes were blocked in 5% skim milk and incubated overnight at 4°C with primary antibodies against LEKTI (29808-1-AP, Proteintech, USA), antibodies against KLK5 (38528, SAB, USA) and HRP-conjugated β-actin recombinant rabbit monoclonal antibody (ET1702-67, HUABIO, China).

Techniques: Expressing, Sequencing, Clinical Proteomics

Journal: The Journal of Experimental Medicine

Article Title: Kallikrein 5 induces atopic dermatitis–like lesions through PAR2-mediated thymic stromal lymphopoietin expression in Netherton syndrome

doi: 10.1084/jem.20082242

Figure Lengend Snippet: KLK5 up-regulates TSLP and proinflammatory molecules in keratinocytes. (A) NHKs were stimulated with 400 nM of recombinant KLK5 for 3 h. TSLP , ICAM1 , IL8 , and TNF -α transcripts are increased by seven-, five-, four-, and fivefold, respectively. No difference was observed for IL-1β , TARC , and MDC ( n = 5). (B) NHK transfected with 60 pmol PAR2 siRNA and stimulated with KLK5 showed PAR2 mRNA down-regulation (by 56%) 4 d after transfection. In PAR2 siRNA-transfected cells, KLK5 is less efficient in inducing TSLP , ICAM1 , IL8 , and TNF-α mRNA (48, 55, 60, and 62% of control, respectively; n = 4). (C) In cells treated with 10 µM NF-κB pathway inhibitor (BAY11-7082), KLK5 efficiency to up-regulate TSLP , ICAM1 , IL8 , and TNF-α mRNA is decreased by 81, 70, 59, and 40%, respectively ( n = 3). For each experiment, the mRNAs of interest were measured by quantitative RT-PCR. Each point is the mean of triplicate amplification for at least three independent experiments ( n ). *, P < 0.05; **, P < 0.01; ***, P < 0.001.

Article Snippet: Cells were transfected with 60 pmol of human PAR2 siRNA or 20 pmol of human KLK5 siRNA (Santa Cruz Biotechnology, Inc.) using the transfection reagent jetSI-ENDO (Ozyme) in serum-free and antibiotic-free EpiLife.

Techniques: Recombinant, Transfection, Control, Quantitative RT-PCR, Amplification

Journal: The Journal of Experimental Medicine

Article Title: Kallikrein 5 induces atopic dermatitis–like lesions through PAR2-mediated thymic stromal lymphopoietin expression in Netherton syndrome

doi: 10.1084/jem.20082242

Figure Lengend Snippet: TSLP and proinflammatory molecules are induced by KLK5 in keratinocytes of NS patients. (A) Quantitative RT-PCR on cultured primary keratinocytes reveals that TSLP is significantly overexpressed by fivefold in three NS patients (NSK) compared with three healthy controls (NHK). ICAM1 , IL8 , TNF-α , TARC , and MDC mRNA are overexpressed in two out of three NSKs but not IL-1β mRNA. Each point represents the mean of two independent experiments for each individual. *, P < 0.05. (B) NHK and NSK were treated with 10 µg/ml brefeldin A, and TSLP was detected by immunofluorescence (green). Blue staining is DAPI. Pictures are representative of two independent brefeldin A treatments, each realized on two NHKs and two NSKs. (C and D) NSKs were transfected with KLK5 siRNA and its efficient down-regulation was confirmed by quantitative RT-PCR, Western blotting (laminin 5 used as loading control), and ELISA, 48 and 72 h after transfection. Results are representative of three independent experiments. (E) KLK5 knockdown induces a decrease of TSLP measured by quantitative RT-PCR. ICAM1 , IL8 , and TNF-α mRNAs were less efficiently reduced. Data are the mean ± SD of one experiment, which is representative of three independent experiments. Bars, 49 µm

Article Snippet: Cells were transfected with 60 pmol of human PAR2 siRNA or 20 pmol of human KLK5 siRNA (Santa Cruz Biotechnology, Inc.) using the transfection reagent jetSI-ENDO (Ozyme) in serum-free and antibiotic-free EpiLife.

Techniques: Quantitative RT-PCR, Cell Culture, Immunofluorescence, Staining, Transfection, Western Blot, Control, Enzyme-linked Immunosorbent Assay, Knockdown

Journal: The Journal of Experimental Medicine

Article Title: Kallikrein 5 induces atopic dermatitis–like lesions through PAR2-mediated thymic stromal lymphopoietin expression in Netherton syndrome

doi: 10.1084/jem.20082242

Figure Lengend Snippet: KLK5 triggers proinflammatory and proallergic microenvironment independently of external stimuli. Lack of KLK5 inhibition by LEKTI initiates proinflammatory and proallergic cascades independently of environmental factors. In LEKTI-deficient keratinocytes, hyperactive KLK5 directly induces TSLP, IL8, and TNF-α overexpression through PAR2 and NF-κB pathway activation. Unregulated KLK5 also degrades desmosomes at the interface between the GR and the SC leading to defective SC adhesion and IL-1β, IL8, and TNF-α secretion by mechanically stressed keratinocytes. In addition, all these cytokines could induce TARC and MDC chemokines secretion by keratinocytes and dermal fibroblasts. These proinflammatory mediators trigger eosinophilic and mast cell recruitment and activation. TSLP has been reported to activate resident LCs, which migrate to draining lymph nodes and promote the differentiation of naive T cells (Th0) into Th2 cells. Collectively, activated keratinocytes together with eosinophilic and mast cells induce pro-Th2 microenvironment favoring the development of an AD-like phenotype.

Article Snippet: Cells were transfected with 60 pmol of human PAR2 siRNA or 20 pmol of human KLK5 siRNA (Santa Cruz Biotechnology, Inc.) using the transfection reagent jetSI-ENDO (Ozyme) in serum-free and antibiotic-free EpiLife.

Techniques: Inhibition, Over Expression, Activation Assay

Journal: Journal of Biological Chemistry

Article Title: Kallikrein-5 Promotes Cleavage of Desmoglein-1 and Loss of Cell-Cell Cohesion in Oral Squamous Cell Carcinoma

doi: 10.1074/jbc.m110.191361

Figure Lengend Snippet: FIGURE 3. Knockdown of KLK-5 expression reduces processing of Dsg1. A, immunocytochemical analysis of KLK5 expression in control (left panel) or KLK5-KD (right panel) SCC25 cells. Cells were cultured on glass coverslips, fixed and incubated with antibodies against KLK5 (1:50 dilution) followed by Al- exa-Fluor-labeled secondary antibodies and counterstained with DAPI. Yellow scale bar, 100 m. B, quantification of immunofluorescent staining with NIH ImageJ for average intensity. C, quantitative real time PCR analysis of KLK5 levels in control and KLK5-KD SCC25 cells. Relative quantification normalized against the housekeeping gene PGK-1 mRNA levels. Graph depicts KLK5 levels in SCC25-KLK5-KD cells normalized relative to SCC25 parental cells (desig- nated as 100). D, analysis of Dsg1 processing in SCC25-KLK5-KD cells. Lysates from duplicate cultures of parental SCC25 cells (lanes 1 and 2) or two clones of SCC25-KLK5-KD cells (lanes 3 and 4) were electrophoresed on 9% SDS-polyacrylamide gels, transferred to PVDF membrane and immunoblotted with anti- Dsg1 (1:1000; upper panel), anti-E-cadherin (1:1000; middle panel), or anti-GAPDH (1:4000; lower panel) followed by peroxidase-conjugated secondary anti- body (1:4000) and peroxidase substrate. Arrow denotes migration position of full-length Dsg1 (165 kDa); arrowhead denotes migration position of cleavage product (130 kDa). E, densitometric quantitation of band density of Dsg1 cleavage product denoted by arrowhead in D. F, densitometric quantitation of band density of corresponding blots shown in D.

Article Snippet: To generate SCC25 cells with reduced KLK5 expression (KLK5 knockdown, designated SCC25-KLK5-KD), pGFPV-RS plasmid containing a gene-specific shRNA sequence silencing KLK5 (GTG TTG GTG CAT CCA CAG TGG CTG CTC AC) (OriGene Technologies, Rockville, MD) was used to transfect SCC25 cells using the Human Keratinocyte Nucleofector kit (Amaxa GmbH, Germany).

Techniques: Knockdown, Expressing, Control, Cell Culture, Incubation, Labeling, Staining, Real-time Polymerase Chain Reaction, Quantitative Proteomics, Clone Assay, Membrane, Migration, Quantitation Assay

Journal: Journal of Biological Chemistry

Article Title: Kallikrein-5 Promotes Cleavage of Desmoglein-1 and Loss of Cell-Cell Cohesion in Oral Squamous Cell Carcinoma

doi: 10.1074/jbc.m110.191361

Figure Lengend Snippet: FIGURE 4. Ultrastructural analysis of desmosomes. SCC25 or SCC25- KLK5-KD cells were grown on coverslips to confluence, then fixed and pro- cessed for transmission electron microscopy. Ultrathin sections were exam- ined with a JEOL 1400 Transmission Electron Microscope. Representative TEM image from (A) SCC25 and (B) SCC25-KLK5-KD cells. Scale bar, 0.2 m. C, quantitation of desmosome number/field from a minimum of 70 images each of SCC25 and SCC25-KLK5-KD cells.

Article Snippet: To generate SCC25 cells with reduced KLK5 expression (KLK5 knockdown, designated SCC25-KLK5-KD), pGFPV-RS plasmid containing a gene-specific shRNA sequence silencing KLK5 (GTG TTG GTG CAT CCA CAG TGG CTG CTC AC) (OriGene Technologies, Rockville, MD) was used to transfect SCC25 cells using the Human Keratinocyte Nucleofector kit (Amaxa GmbH, Germany).

Techniques: Transmission Assay, Electron Microscopy, Microscopy, Quantitation Assay

Journal: Journal of Biological Chemistry

Article Title: Kallikrein-5 Promotes Cleavage of Desmoglein-1 and Loss of Cell-Cell Cohesion in Oral Squamous Cell Carcinoma

doi: 10.1074/jbc.m110.191361

Figure Lengend Snippet: FIGURE 5. Overexpression of KLK-5 expression induces processing of Dsg1. A, immunocytochemical analysis of KLK5 expression in control (left panel) OKF/6 cells or OKF/6 cells transfected with a KLK5 expression vector to generate OKF/6-KLK5 cells (right panel). Cells were cultured on glass coverslips, fixed, and incubated with antibodies against KLK5 (1:50 dilution) followed by Alexa-Fluor-labeled secondary antibodies and counterstained with DAPI. Scale bar 100 m. B, quantification of immunofluorescent staining with NIH ImageJ for average intensity. C, quantitative real time PCR analysis of KLK5 lev- els in control and KLK5 OKF/6 cells. Relative quantification normalized against the housekeeping gene PGK-1 mRNA levels. Graph depicts KLK5 levels in OKF/6-KLK5 cells normalized relative to OKF/6 parental cells (designated as 100). D, analysis of Dsg1 processing in OKF/6-KLK5 cells. Lysates from paren- tal OKF/6 cells (lane 1), vector-transfected OKF/6 cells (lane 2), and OKF/6-KLK5 cells (lane 3) were electrophoresed on 9% SDS-polyacrylamide gels, trans- ferred to PVDF membrane, and immunoblotted with anti-Dsg1 (1:1000; upper panel), anti-E-cadherin (1:1000; middle panel) or anti-GAPDH (1:4000; lower panel) followed by peroxidase-conjugated secondary antibody (1:4000) and peroxidase substrate. The arrow denotes migration position of full-length Dsg1 (165 kDa); arrowhead denotes migration position of cleavage product (130 kDa). E, densitometric quantitation of band density of Dsg1 cleavage product denoted by arrowhead in D. F, densitometric quantitation of band density of corresponding blots shown in D.

Article Snippet: To generate SCC25 cells with reduced KLK5 expression (KLK5 knockdown, designated SCC25-KLK5-KD), pGFPV-RS plasmid containing a gene-specific shRNA sequence silencing KLK5 (GTG TTG GTG CAT CCA CAG TGG CTG CTC AC) (OriGene Technologies, Rockville, MD) was used to transfect SCC25 cells using the Human Keratinocyte Nucleofector kit (Amaxa GmbH, Germany).

Techniques: Over Expression, Expressing, Control, Transfection, Plasmid Preparation, Cell Culture, Incubation, Labeling, Staining, Real-time Polymerase Chain Reaction, Quantitative Proteomics, Membrane, Migration, Quantitation Assay

Journal: Journal of Biological Chemistry

Article Title: Kallikrein-5 Promotes Cleavage of Desmoglein-1 and Loss of Cell-Cell Cohesion in Oral Squamous Cell Carcinoma

doi: 10.1074/jbc.m110.191361

Figure Lengend Snippet: FIGURE 6. Effect of KLK5 expression on cell-cell aggregation dynamics. Single cell suspensions of (A) SCC25, (B) SCC25-KLK5-KD, (C) OKF/6, or (D) OKF/6- KLK5 were incubated in culture medium containing 0.5% BSA and rotated for 7 h. At the indicated time points, aliquots were removed and photographed to visualize cell-cell aggregation. E and H, quantitation of aggregation kinetics. The number of single cells remaining in the suspension at each time point was enumerated and is shown relative to time 0 (100% single cells). (Closed circle) SCC25, (open circle) SCC25-KLK5-KD, (closed triangle) OKF/6, (open triangle) OKF/6-KLK5. F, G, I, J, distribution of cellular aggregates. Within a high-powered field, the number of cellular clusters comprised of (black bar) 10 cells, (white bar) 10–50 cells, and (gray bar) 50 cells was enumerated at the designated time points. (F) SCC25, (G) SCC25-KLK5-KD, (I) OKF/6, (J) OKF/6-KLK5.

Article Snippet: To generate SCC25 cells with reduced KLK5 expression (KLK5 knockdown, designated SCC25-KLK5-KD), pGFPV-RS plasmid containing a gene-specific shRNA sequence silencing KLK5 (GTG TTG GTG CAT CCA CAG TGG CTG CTC AC) (OriGene Technologies, Rockville, MD) was used to transfect SCC25 cells using the Human Keratinocyte Nucleofector kit (Amaxa GmbH, Germany).

Techniques: Expressing, Incubation, Quantitation Assay, Suspension

Journal: Journal of Biological Chemistry

Article Title: Kallikrein-5 Promotes Cleavage of Desmoglein-1 and Loss of Cell-Cell Cohesion in Oral Squamous Cell Carcinoma

doi: 10.1074/jbc.m110.191361

Figure Lengend Snippet: FIGURE 7. Effect of KLK5 expression on monolayer cohesion. Cell-cell adherent monolayers were separated from culture dishes by pulsing with dispase as described under “Experimental Procedures,” transferred to conical tubes affixed to a rocking platform, and subjected to 50 inversion cycles. A, B, D, E, aliquots were photographed to visualize relative monolayer cohesion or dissociation. C and F, total number of fragments present following mechanical dis- ruption was quantified.

Article Snippet: To generate SCC25 cells with reduced KLK5 expression (KLK5 knockdown, designated SCC25-KLK5-KD), pGFPV-RS plasmid containing a gene-specific shRNA sequence silencing KLK5 (GTG TTG GTG CAT CCA CAG TGG CTG CTC AC) (OriGene Technologies, Rockville, MD) was used to transfect SCC25 cells using the Human Keratinocyte Nucleofector kit (Amaxa GmbH, Germany).

Techniques: Expressing

Journal: Cancer Cell International

Article Title: Characterization of KLK5 -high epithelial cells and their interactions with the tumor microenvironment in high-grade serous ovarian cancer

doi: 10.1186/s12935-026-04273-8

Figure Lengend Snippet: Elevated KLK5 expression correlates with poor survival in ovarian cancer. A . Among the up-regulated genes identified across 29 cancer types, 70 genes were specifically overexpressed in ovarian cancer, as determined from the TCGA datasets analyzed using GEPIA2 ( http://gepia2.cancer-pku.cn/#index ). B . KLK5 emerged as the most significantly up-regulated gene (Log 2 FC = 5.759; P-value < 0.001) among the 70 ovarian cancer-specific genes. C . High levels of KLK5 in tumor tissues were significantly associated with reduced disease-free survival. D . KLK5 expression in tumor tissues was notably increased in advanced stages of ovarian cancer (P-value = 0.0455)

Article Snippet: The human qPCR primer pairs GAPDH (Cat No. HP205798 ) and KLK5 (Cat No. HP228990 ) from Origene were used.

Techniques: Expressing

Journal: Cancer Cell International

Article Title: Characterization of KLK5 -high epithelial cells and their interactions with the tumor microenvironment in high-grade serous ovarian cancer

doi: 10.1186/s12935-026-04273-8

Figure Lengend Snippet: The effect of KLK5 on migration, invasion, apoptosis, and proliferation of HO8910 cells. A . The significantly enhanced of migration in the HO8910 cell line with the addition of KLK5 (400ng/mL; P < 0.05). B .The significantly enhanced of invasion in the HO8910 cells with the addition of KLK5 (400 ng/mL in upper chamber; P < 0.05). C . No significantly differences of invasion in the HO8910 cell line with the addition of KLK5 in lower chamber. D . The apoptosis of HO8910 cells treated with or without KLK5 (200 and 400 ng/mL). E , F . The proliferation of HO8910 cells treated with or without KLK5. The line plot in Fig. 2E and F show the mean ± SEM

Article Snippet: The human qPCR primer pairs GAPDH (Cat No. HP205798 ) and KLK5 (Cat No. HP228990 ) from Origene were used.

Techniques: Migration

Journal: Cancer Cell International

Article Title: Characterization of KLK5 -high epithelial cells and their interactions with the tumor microenvironment in high-grade serous ovarian cancer

doi: 10.1186/s12935-026-04273-8

Figure Lengend Snippet: KLK5 is significantly elevated in epithelial cells of high-grade serous ovarian cancer based on single-cell RNA sequencing. A . Uniform Manifold Approximation and Projection (UMAP) visualization of 151,729 cells from 60 samples in publicly available single-cell RNA sequencing datasets from GEO ( https://www.ncbi.nlm.nih.gov/geo/ ). Cells were clustered into seven major cell types, including B cells (B), endothelial cells (Endo), epithelial cells (Epi), fibroblasts (Fib), myeloid cells (Mye), plasma cells (Plasma), and T cells/NK cells (T_NK), based on cell type-specific markers. B . Dot plot illustrating the expression of known marker genes for each cell type. C . UMAP plot displaying the expression levels of KLK5 across the cell clusters. D . KLK5 expression was significantly elevated in epithelial cells of high-grade serous ovarian cancer (HGSOC) compared to normal controls (Log2FC = 0.502, adjusted P < 0.001). E . Spatial transcriptomics shows the expression of KLK5 in HGSOC. OCS, ovarian carcinosarcoma; OCCC, ovarian clear-cell carcinoma; OEC, ovarian endometrioid carcinoma; HGSOC, high-grade serous ovarian cancer; LGSOC, low-grade serous ovarian cancer

Article Snippet: The human qPCR primer pairs GAPDH (Cat No. HP205798 ) and KLK5 (Cat No. HP228990 ) from Origene were used.

Techniques: Single Cell, RNA Sequencing, Clinical Proteomics, Expressing, Marker, Spatial Transcriptomics

Journal: Cancer Cell International

Article Title: Characterization of KLK5 -high epithelial cells and their interactions with the tumor microenvironment in high-grade serous ovarian cancer

doi: 10.1186/s12935-026-04273-8

Figure Lengend Snippet: Cell-cell interaction network between epithelial cells and the other cells. A . Venn diagram illustrating the cell-cell interaction network among different cell types in each subtype of ovarian cancer. B . Specific cell-cell interactions in HGSOC that correspond to up-regulated genes in epithelial cells from HGSOC. C . Corresponding pathway of the specific cell-cell interactions shown in Fig. 4B. D . Expression levels of KLK5 (Log 2 FC = 1.720; adjusted P < 0.001) in KLK5 -high HGSOC, KLK5 -low HGSOC, and normal controls. E. Expression levels of COL1A1 (Log 2 FC = 1.153; adjusted P = 0.0085), COL1A2 (Log 2 FC = 0.728; adjusted P < 0.001), and COL6A2 (Log 2 FC = 0.546; P < 0.001) were significantly elevated in fibroblast cells from the KLK5 -high HGSOC compared to those in KLK5 -low HGSOC or normal controls. *** indicates the P value < 0.001

Article Snippet: The human qPCR primer pairs GAPDH (Cat No. HP205798 ) and KLK5 (Cat No. HP228990 ) from Origene were used.

Techniques: Expressing

Journal: Cancer Cell International

Article Title: Characterization of KLK5 -high epithelial cells and their interactions with the tumor microenvironment in high-grade serous ovarian cancer

doi: 10.1186/s12935-026-04273-8

Figure Lengend Snippet: Transcriptomic changes induced by KLK5 overexpression in ovarian cancer cell lines and the positive relationship between AGRN and KLK5 in single cell RNA datasets of HGSOC. A . Quantitative PCR analysis revealed a significant difference in KLK5 gene expression between the KLK5 -overexpressing SKOV3 cell line (KLK5_OE) and the empty vector control group (Control) ( P < 0.05). B . Transcriptome sequencing analysis identified genes with significant differential expression between KLK5 -overexpressing SKOV3 cells and the empty vector control, where differences with |logFC| > 0.585 and P < 0.05 were considered statistically significant. C . Genes significantly upregulated in KLK5 -overexpressing SKOV3 cells were enriched in the type I interferon synthesis and activation pathways, as indicated by GO enrichment analysis. D . Quantitative PCR analysis revealed a significant difference in KLK5 gene expression between the KLK5 -overexpressing HO-8910 cell line (KLK5_OE) and the empty vector control group (Control) ( P < 0.05). E . Transcriptome sequencing analysis identified genes with significant differential expression between KLK5 -overexpressing HO-8910 cells and the empty vector control, where differences with |logFC| > 0.585 and P < 0.05 were considered statistically significant. F . Venn diagram showing differentially expressed genes (DEGs) between KLK5 -high HGSOC and KLK5 -low HGSOC/normal controls in single cell RNA datasets. G . GO pathway enrichment analysis of common DEGs in epithelial cells from KLK5 -high HGSOC compared to KLK5 -low HGSOC and normal controls, respectively. H . DEGs of epithelial cells between the KLK5 -expression group (Counts ≥ 2) and the null KLK5 -expression group (Counts = 0 in HGSOC. I . KEGG (left) and GO (right) pathway enrichment analyses of DEGs of epithelial cells between the KLK5 -expression group (Counts ≥ 2) and the null KLK5 -expression group (Counts = 0 in HGSOC. J . Positive correlation between KLK5 and AGRN , as demonstrated using the GEPIA2 database ( http://gepia2.cancer-pku.cn/ )

Article Snippet: The human qPCR primer pairs GAPDH (Cat No. HP205798 ) and KLK5 (Cat No. HP228990 ) from Origene were used.

Techniques: Over Expression, Single Cell, Real-time Polymerase Chain Reaction, Gene Expression, Plasmid Preparation, Control, Sequencing, Quantitative Proteomics, Activation Assay, Expressing

Journal: Cancer Cell International

Article Title: Characterization of KLK5 -high epithelial cells and their interactions with the tumor microenvironment in high-grade serous ovarian cancer

doi: 10.1186/s12935-026-04273-8

Figure Lengend Snippet: Enhanced tumor-associated macrophages in KLK5-high HGSOC. A. Immune cell infiltration profiles of TCGA-KLK5 High , TCGA-KLK5 Low and normal controls, analyzed using the ssGSEA algorithm. B . Subtypes and their proportions of monocytes and macrophages. C . Unsupervised trajectory analysis illustrating state transitions between monocytes and macrophages. The branched trajectory is color-coded by cell states and cell subtypes. D . GO terms enriched from up-regulated genes in macrophage subtypes. E . Differentially expressed genes (DEGs) in THP-1-derived macrophages after co-culture with KLK5-overexpressing (versus empty vector control) HO-8910 cells. (|log₂FC| > 0.585 and P < 0.05). F . Differentially expressed genes (DEGs) in THP-1-derived macrophages after co-culture with KLK5-overexpressing (versus empty vector control) SKOV3 cells. (|log₂FC| > 0.585 and P < 0.05)

Article Snippet: The human qPCR primer pairs GAPDH (Cat No. HP205798 ) and KLK5 (Cat No. HP228990 ) from Origene were used.

Techniques: Derivative Assay, Co-Culture Assay, Plasmid Preparation, Control

Journal: Cancer Cell International

Article Title: Characterization of KLK5 -high epithelial cells and their interactions with the tumor microenvironment in high-grade serous ovarian cancer

doi: 10.1186/s12935-026-04273-8

Figure Lengend Snippet: Immune cell-cell interactions in KLK5 -high HGSOC. A . Uniform Manifold Approximation and Projection (UMAP) plot of T and NK cells, color-coded by cell subtypes. B . Relative proportions of T and NK cell subsets in each group. C . GO terms enriched from up-regulated genes in T_CD8_Cytotoxic cells. D . GO terms enriched from up-regulated genes in NK_XCL1 cells. E . Unique Cell-cell interactions between myeloid cells (Mye) and T-NK cells in KLK5 -high HGSOC

Article Snippet: The human qPCR primer pairs GAPDH (Cat No. HP205798 ) and KLK5 (Cat No. HP228990 ) from Origene were used.

Techniques:

Journal: Dermatology and Therapy

Article Title: Topical Treatment of Rosacea with Ivermectin Inhibits Gene Expression of Cathelicidin Innate Immune Mediators, LL-37 and KLK5, in Reconstructed and Ex Vivo Skin Models

doi: 10.1007/s13555-017-0176-3

Figure Lengend Snippet: Ivermectin treatment inhibits KLK5 secretion from human epidermal cells. a NHEK cells were pre-treated with ivermectin, azelaic acid, metronidazole at 1 µM or vehicle control overnight before 48 h stimulation with 100 nM calcitriol. KLK5 release was measured by ELISA. Data show mean ± SD from 3 independent experiments. b RHE was pre-treated with ivermectin, azelaic acid, metronidazole at 1% w/v or vehicle control (100% ethanol) overnight before 24 h topical stimulation with 10 µM calcitriol. KLK5 release was measured by ELISA. Data show mean ± SD from two independent experiments. c Ex vivo skin biopsies were pre-treated with ivermectin, azelaic acid, metronidazole at 1% w/v or vehicle control overnight before 48 h topical stimulation with 10 µM calcitriol. KLK5 release was measured by ELISA. Data show mean ± SD from two independent experiments. NS not significant. ** P < .01, *** P < .001, Student’s t test. d Immunohistologic staining of paraffin-embedded reconstructed epidermis using anti-KLK5 antibody. Top panel RHE was pre-treated with vehicle control overnight and then was stimulated with vehicle control. Middle panel RHE was pre-treated with vehicle control overnight before being stimulated topically with 10 µM calcitriol. Lower panel RHE was pre-treated with ivermectin at 1% w/v overnight before being stimulated topically with 10 µM calcitriol. The epidermis was then paraffin-embedded and stained with anti-KLK5 antibody (in red ). Scale bars 100 µm

Article Snippet: Immunohistochemistry was performed on Discovery Xt, Ventana (Roche) using a rabbit anti-KLK5 antibody (TA321822; OriGene Technologies, Rockville, MD, USA) and a mouse anti-LL-37 antibody (sc-166770 clone D-5; Santa Cruz Biotechnology, Dallas, TX, USA).

Techniques: Control, Enzyme-linked Immunosorbent Assay, Ex Vivo, Staining

Journal: Dermatology and Therapy

Article Title: Topical Treatment of Rosacea with Ivermectin Inhibits Gene Expression of Cathelicidin Innate Immune Mediators, LL-37 and KLK5, in Reconstructed and Ex Vivo Skin Models

doi: 10.1007/s13555-017-0176-3

Figure Lengend Snippet: Ivermectin treatment inhibits cathelicidin pathway gene expression in ex vivo human skin biopsies. Ex vivo skin biopsies were pre-treated with ivermectin at 1% w/v or vehicle control (100% ethanol) before either 24- or 72-h topical stimulation with 10 µM calcitriol. Tissues were lysed and RNA extracted in order to perform qRT-PCR using pre-developed Taqman Assay probes. a Graphical representation show fold change modulation of CAMP, KLK8, KLK7 and KLK5 gene expression after 24-h stimulation and 72-h stimulation. b Table represents normalized Ct values considering housekeeping gene expression (GAPDH, HPRT1 and GUSB) of control sample (vehicle-treated) at 24 or 72 h. The lower is the normalized Ct value; the higher is the gene expression value. Fold induction are calculated for calcitriol-treated sample using vehicle as reference sample. Data show mean from three independent experiments. Analyses were performed using Array Studio software. Results were considered significant when at least a twofold difference in expression level was detected and statistical analysis revealed an adjusted P value (FDR_BH) <0.05

Article Snippet: Immunohistochemistry was performed on Discovery Xt, Ventana (Roche) using a rabbit anti-KLK5 antibody (TA321822; OriGene Technologies, Rockville, MD, USA) and a mouse anti-LL-37 antibody (sc-166770 clone D-5; Santa Cruz Biotechnology, Dallas, TX, USA).

Techniques: Gene Expression, Ex Vivo, Control, Quantitative RT-PCR, TaqMan Assay, Software, Expressing

Journal: NPJ Biofilms and Microbiomes

Article Title: Nasal symbiont Staphylococcus epidermidis restricts the cellular entry of influenza virus into the nasal epithelium

doi: 10.1038/s41522-022-00290-3

Figure Lengend Snippet: A Scatterplots indicating enriched genes related with serine-type peptidase. B PPI network of DEGs linked with serine-type peptidase indicates the protein – protein interaction with Serpine1 and Serpine2 (PP protein – protein). C Heatmap depicting the expression levels of genes related with serine-type peptidase inhibitor activity differentially expressed in S. epidermidis– inoculated NHNE cells depending on a cellular subset. D Serpine1 mRNA and E protein levels were monitored by real-time PCR and western blot analysis following S. epidermidis inoculation at 1 dpi with transfection with cont shRNA and uPA shRNA. F Serpine1 (blue bar) and PLAU mRNAs (red bar) were compared in NHNE cells after S. epidermidis inoculation. G ELISA results showed the secreted protein levels of Serpine1 (blue bar) and uPA (red bar) in S. epidermidis -inoculated NHNE cells. H The intracellular protein levels of HAT and Serpine1 were measured in the cell lysate of S. epidermidis– inoculated NHNE cells at 4, 8, and 24 h using western blot analysis. I The mRNA levels of proteases, such as TMPRSS2, TMPRSS11E, TMPRSS11F, and KLK5, which were targeted by Serpine1, were measured using the cell lysate of S. epidermidis -inoculated NHNE cells at 4, 8, and 24 h. Western blot results are representative of five independent experiments. Real-time PCR and plaque assay results are presented as mean ± SD values from five independent experiments. * p < 0.05 vs. mock-infected NHNE cells.

Article Snippet: Primers for human or mouse Serpine1 (assay ID Hs00167155_m1, Mm00435858_m1), Serpine2 (assay ID Hs00299953_m1, Mm00436753_m1), TMPRSS11E (assay ID Hs01070171_m1, Mm01212186_m1), TMPRSS11F (assay ID Hs01592083_m1, Mm00812591_m1), TMPRSS2 (assay ID Hs01122322_m1, Mm00443687_m1), and KLK5 (assay ID Hs01548153_m1, Mm01203811_m1) were purchased from Applied Biosystems (Foster City, CA, USA).

Techniques: Expressing, Activity Assay, Real-time Polymerase Chain Reaction, Western Blot, Transfection, shRNA, Enzyme-linked Immunosorbent Assay, Plaque Assay, Infection