Journal: PLoS ONE

Article Title: SERPINE2 Inhibits IL-1α-Induced MMP-13 Expression in Human Chondrocytes: Involvement of ERK/NF-κB/AP-1 Pathways

doi: 10.1371/journal.pone.0135979

Figure Lengend Snippet: A, B . Human SERPINE2 mRNA expression after interleukin IL-1α treatment (0.05–10 ng/mL) during 24 hours in human primary chondrocytes and in T/C-28a2 chondrogenic cells. C, D . Representative western blot of human SERPINE2 protein expression in lysates obtained from human primary chondrocytes and T/C-28a2 chondrogenic cells treated with interleukin IL-1α (0.05–10 ng/mL) for 24 h. β-actin was used to ensure equal sample loading. Data are means ± S.E.M. of at least 3 independent experiments. *P<0.05, **P<0.01 and ***P<0.001 vs untreated control cells.

Article Snippet: Blots were incubated with the appropriate antibody: anti-human SERPINE2 (R&D, MN, USA); anti-human MMP-13 (Santa Cruz, CA, USA); anti-phospho ERK1/2 (Millipore, MA, USA); anti-ERK1/2 (Millipore, MA, USA), anti-p65 (Santa Cruz, CA,USA); anti-c-jun (Santa Cruz, CA, USA).

Techniques: Expressing, Western Blot

Journal: PLoS ONE

Article Title: SERPINE2 Inhibits IL-1α-Induced MMP-13 Expression in Human Chondrocytes: Involvement of ERK/NF-κB/AP-1 Pathways

doi: 10.1371/journal.pone.0135979

Figure Lengend Snippet: A. Human MMP-13 mRNA expression in T/C-28a2 chondrocytes incubated with SERPINE2 (0.4 ng/mL) in presence or not of IL-1α (0.5 ng/mL) for 24 h. B. Representative western blot of human MMP-13 protein expression in lysates obtained from T/C-28a2 chondrogenic cells treated with with SERPINE2 (0.4 ng/mL) in presence or not of IL-1α (0.5 ng/mL) for 24 h. β-actin was used to ensure equal sample loading. Low panel. Densitometric analysis of at least three independent experiments. C. Human MMP-13 mRNA expression in human primary chondrocytes incubated with SERPINE2 (0.4 ng/mL) in presence or not of IL-1α (0.5 ng/mL) for 24 h. Data are means ± S.E.M. of at least 3 independent experiments. **P<0.01 and ***P<0.001 vs untreated control cells; ## P<0.01 and ### P<0.001 vs IL-1α-stimulated chondrocytes.

Article Snippet: Blots were incubated with the appropriate antibody: anti-human SERPINE2 (R&D, MN, USA); anti-human MMP-13 (Santa Cruz, CA, USA); anti-phospho ERK1/2 (Millipore, MA, USA); anti-ERK1/2 (Millipore, MA, USA), anti-p65 (Santa Cruz, CA,USA); anti-c-jun (Santa Cruz, CA, USA).

Techniques: Expressing, Incubation, Western Blot

Journal: Oncotarget

Article Title: Serpin E2 promotes breast cancer metastasis by remodeling the tumor matrix and polarizing tumor associated macrophages

doi: 10.18632/oncotarget.12927

Figure Lengend Snippet: ( A ) Masson trichrome staining performed on 4T1 Ctrl and shSerpine2 tumors. Representative images show increased collagen encapsulation (blue) of SerpinE2 KD tumors. Scale bar100μm. ( B - C ) Intravital imaging using (IVI-MP) of 4T1 Ctrl and shSerpine2 tumors. (B) Representative images show collagen I fibers detected by the SHG signal (cyan) at the surface of 4T1 tumors; scale bar25μm. (C) Average SHG signal intensity was determined per 100μm z-stack; data are acquired from 19-30 separate z-stacks from 3 mice per cell line. * P < 0.00036. D. IVI-MP performed on mice bearing GFP-labeled 4T1 control and shSerpinE2 tumors. Representative images show GFP-labeled tumor cells (green), phagocytic dextran positive cells (red); SHG imaging identified collagen I fibers (cyan). Scale bars25μm. (E-F) ( E ) GFP-labeled 4T1 tumor-bearing mice were treated with control liposomes or clodronate-containing liposomes until IVI-MP was performed. Representative images are shown as in (D). ( F ) Quantification of SHG (cyan) signal intensity in 100 μm Z-stacks of tumors in treated animals. Data are mean ± SEM of measurements from 40-61 Z-stacks from at least 3 different tumors for each treatment group. * P < 0.016. All data are mean ± SEM.

Article Snippet: Anti-human SerpinE2 Ab (MAB2980) was from R&D systems.

Techniques: Staining, Encapsulation, Imaging, Labeling, Control, Liposomes

Journal: Oncotarget

Article Title: Serpin E2 promotes breast cancer metastasis by remodeling the tumor matrix and polarizing tumor associated macrophages

doi: 10.18632/oncotarget.12927

Figure Lengend Snippet: Conditioned medium (CM) from 4T1 and 168FARN ( A ) or MDA-MB435 ( B ) cultures was incubated overnight with Ab11, then bound serpinE2 was pulled-down with Protein G, and analyzed by western blot with the rodent-specific antibody, 4B3 (A), or a human-specific antibody (B). (A) The right panel shows a short exposure of serpinE2 complexes from 4T1 CM, but none in 168FARN CM (long exposure on the left), as well as an IP of the preformed tPA/serpinE2 complex. ( C ) SFM loaded with purified human serpinE2 or the preformed serpinE2/tPA complex was IP'd with Ab11 as in (A) & (B) and analyzed by western blot with a human serpinE2-specific antibody. The serpinE2 complex and uncomplexed serpinE2 are indicated with arrowheads.

Article Snippet: Anti-human SerpinE2 Ab (MAB2980) was from R&D systems.

Techniques: Incubation, Western Blot, Purification

Journal: Oncotarget

Article Title: Serpin E2 promotes breast cancer metastasis by remodeling the tumor matrix and polarizing tumor associated macrophages

doi: 10.18632/oncotarget.12927

Figure Lengend Snippet: The LRP1 receptor is expressed on tumor cells and on macrophages and we propose that serpinE2 targeting with Ab11 impacts on LRP1 signaling in both cell types. ( A ) In metastatic tumors, serpinE2-LRP1 binding stimulates ERK signaling and secretion of CCL2. Tumor cells display hyperactivation of receptor tyrosine kinases (RTKs); FGFRs, which are active in the 4T1 model, also stimulate ERK pathway activation. We speculate that in macrophages, the combination of high CCL2 levels and serpinE2-activated LRP1 promotes the M2 phenotype. Indeed, there are high levels of phagocytic, Texas-red positive M2 tumor associated macrophages (TAMs) in the metastatic tumors; they are known to be responsible for degrading the matrix during tumor development. ( B ) Ab11 or sepinE2 KD (not drawn in model) lowers ERK pathway activity and the secretion of CCL2, and stimulates TIMP1 secretion. Tyrosine kinase inhibitors (TKIs) block RTK signaling. The matrix-degrading M2 TAMs are decreased in Ab11-treated tumors. Moreover, depleting macrophages with clodronate liposomes results in deposition of a dense collagen matrix, similar to that observed in Ab11-treated tumors. Thus, we propose that the drop in CCL2, which contributes to a decrease in M2 TAMs, as well as blocking serpinE2, which skews the TAMs towards the M1 phenotype, causes the emergence of a dense collagen matrix that inhibits intravasation and metastatic dissemination.

Article Snippet: Anti-human SerpinE2 Ab (MAB2980) was from R&D systems.

Techniques: Binding Assay, Activation Assay, Activity Assay, Blocking Assay, Liposomes

Journal: Oncogene

Article Title: A slow-cycling subpopulation of melanoma cells with highly invasive properties

doi: 10.1038/onc.2017.341

Figure Lengend Snippet: Label-retaining, disseminated melanoma cells express high levels of SerpinE2. ( a ) Table lists proteins identified after whole-cell proteomic analyses of LRC and non-LRC. The last column lists the cell lines with differential expression of the indicated protein. ( b ) Violin plots (left panels) illustrate quantification of SerpinE2 (white spot, top) and BMP1 (white spots, bottom) as determined by mRNA FISH. mRNA counts for single cells are shown on the Y -axes. Note differences in Y -axes. Representative photomicrographs are shown on the right (total magnification: 100 ×). ( c ) Protein expression was analyzed by immunofluorescence and quantified at single-cell levels (scatter plots on left and representative images on right). LRC (red), non-LRC (orange), total cells (gray). ( d ) Flow cytometry of disseminated WM989 melanoma cells detected in three mouse organs (top panel). The percentage of cells double positive for CD146 and SerpinE2 is shown in the upper right quadrants; the lower right quadrants indicate the percentage of cells singly positive for CD146. Micrographs show examples of disseminated SerpinE2-positive cells (arrows, middle and right panels) in mouse lungs. Left panel: negative control.

Article Snippet: Samples were boiled 10 min in NuPAGE LDS sample buffer (Thermo Fisher Scientific), loaded on 10% gels, transferred onto nitrocellulose filters and immunoblotted with mouse anti-human SerpinE2 antibody (R&D), diluted 1:500.

Techniques: Expressing, Immunofluorescence, Flow Cytometry, Negative Control

Journal: Oncogene

Article Title: A slow-cycling subpopulation of melanoma cells with highly invasive properties

doi: 10.1038/onc.2017.341

Figure Lengend Snippet: SerpinE2 drives melanoma invasiveness. ( a ) Boyden chamber invasion assay of sorted label-retaining cells (LRC) or non-LRC in presence (blue columns) or absence (gray columns) of human recombinant SerpinE2. Bars show fold change in invasion (mean±s.e.m.) of samples with SerpinE2 over untreated cells. ( b ) Invasion assay in presence of an anti-human SerpinE2 neutralizing antibody or Isotype control (ctrl). The percentage of invading cells after neutralization is shown. Data represent mean±s.e.m. of three independent experiments. ( c ) Invasion assay after SerpinE2 knockdown. Data shown are percentage of invading cells found after silencing with 5 different shRNA (Sh_13-17) relative to Sh_ctrl (ctrl). Bars represent mean±s.e.m.

Article Snippet: Samples were boiled 10 min in NuPAGE LDS sample buffer (Thermo Fisher Scientific), loaded on 10% gels, transferred onto nitrocellulose filters and immunoblotted with mouse anti-human SerpinE2 antibody (R&D), diluted 1:500.

Techniques: Invasion Assay, Recombinant, Neutralization, shRNA

Journal: Oncogene

Article Title: A slow-cycling subpopulation of melanoma cells with highly invasive properties

doi: 10.1038/onc.2017.341

Figure Lengend Snippet: SerpinE2 expression is increased in malignant cells and correlates with tumor progression. ( a ) Western blot analysis of SerpinE2 secretion by melanoma cells ( n =21) and human melanocytes ( n =5) into culture supernatants. Recombinant SerpinE2 protein (500 ng) was used as a positive control and Ponceau’s staining is shown for loading control. Graph shows quantification (right panel). ( b ) SerpinE2 staining of 3D skin reconstructs with melanocytes (left) and melanoma cells (right). ( c ) GEM_1375 data set analysis of SerpinE2 mRNA expression in melanomas, non-malignant nevi and normal skin. ( d ) Examples of immune histochemistry at two different magnifications of: normal skin, benign nevi, in situ , vertical growth phase (VGP) and lymph node metastatic melanomas ( n =15 for each group). SerpinE2 protein expression is indicated by purple staining. Arrows indicate the dotted SerpinE2 expression pattern found in benign nevi only. ( e ) Intensity score of SerpinE2 expression in normal skin, benign nevi and malignant melanomas (radial growth phase (RGP), VGP and metastatic). Bars represent mean±SEM of tissue sections of 15 specimens from each group; P- value after t -student test are given.

Article Snippet: Samples were boiled 10 min in NuPAGE LDS sample buffer (Thermo Fisher Scientific), loaded on 10% gels, transferred onto nitrocellulose filters and immunoblotted with mouse anti-human SerpinE2 antibody (R&D), diluted 1:500.

Techniques: Expressing, Western Blot, Recombinant, Positive Control, Staining, In Situ

Journal: Frontiers in Oncology

Article Title: SerpinE2 promotes M2 polarization in macrophage to accelerate colorectal cancer progression

doi: 10.3389/fonc.2025.1585935

Figure Lengend Snippet: SERPINE2 expression is significantly upregulated in COAD. (A) Box plot of SERPINE family genes mRNA expression.in COAD(p < 0.05). (B) Box plot of SERPINE1 and SERPINE2 protein expression in COAD(p < 0.05). (C) Box plot of SERPINE2 mRNA expression among groups of different tumor grades in COAD (p < 0.05).

Article Snippet: Concentrations of SERPINE2 (human) in the culture supernatant of treated cells were measured with the use of a commercially available kit (CUSABIO).Seed HCT-8 cells in a 100-mm culture dish at a density of 1 × 106 cells in complete medium (RPMI-1640,+10%FBS).At 60–80% confluency, perform siRNA transfection using [Lipofectamine 3000].

Techniques: Expressing

Journal: Frontiers in Oncology

Article Title: SerpinE2 promotes M2 polarization in macrophage to accelerate colorectal cancer progression

doi: 10.3389/fonc.2025.1585935

Figure Lengend Snippet: SERPINE2 expression correlates with tumor immunoregulation, division and proliferation. (A) Volcano map of SERPINE2 co-expressed genes in COAD. (B, C) GO and KEGG enrichment analysis of SERPINE2 co-expressed genes obtained from the cBioPortal, the terms p and q < 0.05 were considered to be significantly enriched. Only 20 leading gene sets are displayed in the plot.

Article Snippet: Concentrations of SERPINE2 (human) in the culture supernatant of treated cells were measured with the use of a commercially available kit (CUSABIO).Seed HCT-8 cells in a 100-mm culture dish at a density of 1 × 106 cells in complete medium (RPMI-1640,+10%FBS).At 60–80% confluency, perform siRNA transfection using [Lipofectamine 3000].

Techniques: Expressing

Journal: Frontiers in Oncology

Article Title: SerpinE2 promotes M2 polarization in macrophage to accelerate colorectal cancer progression

doi: 10.3389/fonc.2025.1585935

Figure Lengend Snippet: SERPINE2 is positively correlated with M2 macrophage infiltration in COAD. (A) Scatter plot showing the correlation of 6 different immune cell types and SERPINE2 expression (p < 0.05). The black line in each plot is a fitted linear model indicating the proportion of tropism of the immune cell along with SERPINE2 expression, and the Pearson coefficient was used for the correlation test. (B) Scatter plot showing the correlation of CD68+ Marcophage and SERPINE2 expression (p < 0.05). C.Scatter plot showing the correlation of M1/M2 macrophage and SERPINE2 expression (p < 0.05). The blue line in each plot is a fitted linear model indicating the proportion of tropism of the immune cell along with SERPINE2 expression, and the Pearson coefficient was used for the correlation test. (p < 0.05).

Article Snippet: Concentrations of SERPINE2 (human) in the culture supernatant of treated cells were measured with the use of a commercially available kit (CUSABIO).Seed HCT-8 cells in a 100-mm culture dish at a density of 1 × 106 cells in complete medium (RPMI-1640,+10%FBS).At 60–80% confluency, perform siRNA transfection using [Lipofectamine 3000].

Techniques: Expressing

Journal: Frontiers in Oncology

Article Title: SerpinE2 promotes M2 polarization in macrophage to accelerate colorectal cancer progression

doi: 10.3389/fonc.2025.1585935

Figure Lengend Snippet: SERPINE2 is associated with infiltration of CD68 macrophage in COAD patients. (A) The expression of SEPRINE2 in patient tissues was detected by IHC. Adjacent tissue not expressing SERPINE2; tumor tissue strongly expressing SERPINE2. (B) Scatterplot showing the correlation between SERPINE2 and CD68 expression (p < 0.05). The blue line in each plot is a fitted linear model indicating the relationship between SERPINE2 and CD68 expression. (C) The expression of SERPINE2 and CD68 in patient tissues were detected by IHC. High levels of SERPINE2 expression are associated with increased expression of CD68. (* p<0.05, **p<0.01, *** p<0.001).

Article Snippet: Concentrations of SERPINE2 (human) in the culture supernatant of treated cells were measured with the use of a commercially available kit (CUSABIO).Seed HCT-8 cells in a 100-mm culture dish at a density of 1 × 106 cells in complete medium (RPMI-1640,+10%FBS).At 60–80% confluency, perform siRNA transfection using [Lipofectamine 3000].

Techniques: Expressing

Journal: Frontiers in Oncology

Article Title: SerpinE2 promotes M2 polarization in macrophage to accelerate colorectal cancer progression

doi: 10.3389/fonc.2025.1585935

Figure Lengend Snippet: The external secretion of SERPINE2 from colorectal cancer cells promotes macrophage polarization to accelerate the development of colorectal cancer. (A) Schema diagram showing that THP-1 cells were treated with PMA, followed by the treatment of SERPINE2 recombinant protein as indicated treatments to obtain activated macrophages. Then tumor cells cocultured with these macrophages. were used to subsequent experiments. (B, C) RT-qPCR and Westen blot displayed SERPINE2 mRNA and Protein expression were upregulated in colorectal cancer cells. (D) The secretion of SERPINE2 in colorectal cancer cells was detected by ELISA. (E) The effect of SERPINE2 recombinant protein on macrophage polarization. (F) RT-qPCR displayed conditioned medium (CM) derived from colorectal cancer cells promotes the upregulation of macrophage M2 markers, compared to normal Intestinal cells. (G) ELISA analysis demonstrated that SERPINE2 secretion was significantly reduced after SERPINE2 knockdown. (H) RT-qPCR displayed conditioned medium with SERPINE2 knockdown down-regulated macrophage M2 markers. (I, J) The conditioned medium from macrophages treated with SERPINE2 recombinant protein promoted cancer cell proliferation (measured by CCK-8 assay) and migration (assessed by Transwell assay. (*p<0.05, **p<0.01, ***p<0.001).

Article Snippet: Concentrations of SERPINE2 (human) in the culture supernatant of treated cells were measured with the use of a commercially available kit (CUSABIO).Seed HCT-8 cells in a 100-mm culture dish at a density of 1 × 106 cells in complete medium (RPMI-1640,+10%FBS).At 60–80% confluency, perform siRNA transfection using [Lipofectamine 3000].

Techniques: Recombinant, Quantitative RT-PCR, Expressing, Enzyme-linked Immunosorbent Assay, Derivative Assay, Knockdown, CCK-8 Assay, Migration, Transwell Assay

Journal: Reproductive Biology and Endocrinology : RB&E

Article Title: SERPINE2, an inhibitor of plasminogen activators, is highly expressed in the human endometrium during the secretory phase

doi: 10.1186/1477-7827-9-38

Figure Lengend Snippet: Antibody specificity and presence of the SERPINE2 protein in human uterine fluid . (A) Four hundred nanograms of recombinant human SERPINE2 was resolved on 10% SDS-PAGE and followed by Western blotting using anti-mouse SERPINE2 antiserum (lane 1), an anti-human SERPINE2 antibody (R&D) (lane2), or another anti-human SERPINE2 antibody (Abnova) (lane 3). (B) One hundred micrograms of the extract of endometrial curettage was analyzed by anti-mouse SERPINE2 antiserum (lanes 1 and 2) and an anti-human SERPINE2 antibody (R&D) (lanes 3 and 4). (C) Fifty micrograms of uterine fluid proteins collected from each individual patient ( n = 7) was Western-blotted using anti-mouse SERPINE2 antiserum (1:3000) (upper panel). EP, MP, and LP indicate early-, mid-, and late-proliferative phases, and ES and MS indicate early- and mid-secretory phases, respectively. The blot was also overexposed to clearly display the staining signal (lower panel).

Article Snippet: Membranes were blocked with 10% (w/v) skim milk in phosphate-buffered saline (PBS) (blocking solution) for 2 h, and then incubated with our homemade anti-mouse SERPINE2 antiserum (1: 5000) [ , ], anti-human SERPINE2 antibody (1: 1000, catalog no. AF2980, R&D Systems, Minneapolis, MN, USA), or another anti-human SERPINE2 antibody (1: 1000, product no. H00005270-B01, Abnova) in blocking solution for 2 h at 37°C.

Techniques: Recombinant, SDS Page, Western Blot, Staining

Journal: Reproductive Biology and Endocrinology : RB&E

Article Title: SERPINE2, an inhibitor of plasminogen activators, is highly expressed in the human endometrium during the secretory phase

doi: 10.1186/1477-7827-9-38

Figure Lengend Snippet: Localization of SERPINE2 in the human uterus . Longitudinal sections of the early secretory phase uterus (n = 5) on the slide were incubated with anti-mouse SERPINE2 antiserum and then treated with biotin-conjugated goat-anti-rabbit IgG and HRP-conjugated streptavidin (brown). For contrast, specimens were further stained with hematoxylin (blue). The representative picture is shown. Magnified pictures of the luminal epithelium (A), glandular epithelium (B), and myometrium (C) are shown. Bar = 50 μm. bv, blood vessel; ge, glandular epithelium; le, luminal epithelium; m, muscle; s, stroma.

Article Snippet: Membranes were blocked with 10% (w/v) skim milk in phosphate-buffered saline (PBS) (blocking solution) for 2 h, and then incubated with our homemade anti-mouse SERPINE2 antiserum (1: 5000) [ , ], anti-human SERPINE2 antibody (1: 1000, catalog no. AF2980, R&D Systems, Minneapolis, MN, USA), or another anti-human SERPINE2 antibody (1: 1000, product no. H00005270-B01, Abnova) in blocking solution for 2 h at 37°C.

Techniques: Incubation, Staining

Journal: Reproductive Biology and Endocrinology : RB&E

Article Title: SERPINE2, an inhibitor of plasminogen activators, is highly expressed in the human endometrium during the secretory phase

doi: 10.1186/1477-7827-9-38

Figure Lengend Snippet: Glandular epithelial expression of the SERPINE2 protein in the endometrium during the menstrual cycle . Sections prepared from endometrial curettage during early-proliferative (EP), mid-proliferative (MP), late-proliferative (LP), early-secretory (ES), mid-secretory (MS), and late-secretory (LS) phases were incubated with anti-mouse SERPINE2 antiserum and then treated as described in Figure 2. Bar = 50 μm. bv, blood vessel; ge, glandular epithelium; s, stroma.

Article Snippet: Membranes were blocked with 10% (w/v) skim milk in phosphate-buffered saline (PBS) (blocking solution) for 2 h, and then incubated with our homemade anti-mouse SERPINE2 antiserum (1: 5000) [ , ], anti-human SERPINE2 antibody (1: 1000, catalog no. AF2980, R&D Systems, Minneapolis, MN, USA), or another anti-human SERPINE2 antibody (1: 1000, product no. H00005270-B01, Abnova) in blocking solution for 2 h at 37°C.

Techniques: Expressing, Incubation

Journal: Reproductive Biology and Endocrinology : RB&E

Article Title: SERPINE2, an inhibitor of plasminogen activators, is highly expressed in the human endometrium during the secretory phase

doi: 10.1186/1477-7827-9-38

Figure Lengend Snippet: Quantification of SERPINE2 protein expression levels in endometrial glands . Representative samples were analyzed by automated cell acquisition and quantification software (A). The expression signal of a respective glandular gland was quantified using HistoQuest software and is presented as a scattergram. Each spot on the scattergram stands for the intensity of one cell (B). The relative SERPINE2 protein expression levels in patients' glandular glands at various sub-phases of the menstrual cycle are shown as bar diagrams (C). Differences are significant among patients at various groups (χ = 69.32, p < 0.0001).

Article Snippet: Membranes were blocked with 10% (w/v) skim milk in phosphate-buffered saline (PBS) (blocking solution) for 2 h, and then incubated with our homemade anti-mouse SERPINE2 antiserum (1: 5000) [ , ], anti-human SERPINE2 antibody (1: 1000, catalog no. AF2980, R&D Systems, Minneapolis, MN, USA), or another anti-human SERPINE2 antibody (1: 1000, product no. H00005270-B01, Abnova) in blocking solution for 2 h at 37°C.

Techniques: Expressing, Software