Journal: Communications Biology

Article Title: Integrated methylome analysis identifies an epigenetically silenced tumor suppressor RASA4 in small cell lung cancer

doi: 10.1038/s42003-025-09440-7

Figure Lengend Snippet: A Volcano plot of differentially expressed genes in SBC-2 cells with stable RASA4 knockdown, relative to those transfected with scramble shRNA. B qRT-PCR shows that SERPINE2 mRNA levels were significantly lower in DMS114 and SHP77 cells with stable over-expression of RASA4, compared to cells stably transfected with an empty vector. SERPINE2 mRNA levels were significantly higher in H446 and SBC-2 cells with stable knockdown of RASA4, compared to cells stably transfected with scramble shRNA. Data are shown as mean ± SD from three independent biological replicates ( n = 3, unpaired Student’s t -test). C ELISA analysis shows that H446 and SBC-2 cells with RASA4 knockdown secreted more SERPINE2 protein to cell culture medium, compared to cells stably transfected with scramble shRNA. Data are shown as mean ± SD from three independent biological replicates ( n = 3, unpaired Student’s t -test). D Immunofluorescence staining of SERPINE2 protein in SBC-2 and H446 cells with stable transfections of scramble shRNA and RASA4 shRNA. (scale bar = 10 μm for SBC-2 cells, scale bar = 20 μm for H446 cells). E Co-IP analysis of protein interaction between RASA4 and SERPINE2 in DMS114 cells with RASA4 stable over-expression. Cell lysates from DMS114 cells with stable transfection of FLAG-tagged RASA4 were immunoprecipitated with anti-FLAG antibody, followed by western blot with anti-SERPINE2 antibody. IgG was used as a negative control. F Western blot shows that the protein expression of SERPINE2 and EMT markers (N-cadherin, E-cadherin, Vinmintin, and snail) following RASA4 stable knockdown and overexpression in SBC-2 and DMS114 cells. G qRT-PCR analysis of stemness markers NANOG , OCT4 , SOX2, and MYC in SBC-2 and DMS114 cells with RASA4 stable knockdown and overexpression. Data are shown as mean ± SD from three independent biological replicates ( n = 3, unpaired Student’s t -test). H Sphere formation assay in SBC-2 and H446 cells with RASA4 stable knockdown, and in DMS114 and SHP77 cells with RASA4 stable overexpression. (scale bar = 50 μm).

Article Snippet: SERPINE2 concentration in cell culture medium was quantified using the Human Glia-Derived Nexin (SERPINE2) ELISA Kit (#CSB-EL021082HU, Cusabio, Wuhan, China) following the manufacturer’s protocol.

Techniques: Knockdown, Transfection, shRNA, Quantitative RT-PCR, Over Expression, Stable Transfection, Plasmid Preparation, Enzyme-linked Immunosorbent Assay, Cell Culture, Immunofluorescence, Staining, Co-Immunoprecipitation Assay, Immunoprecipitation, Western Blot, Negative Control, Expressing, Tube Formation Assay

Journal: Communications Biology

Article Title: Integrated methylome analysis identifies an epigenetically silenced tumor suppressor RASA4 in small cell lung cancer

doi: 10.1038/s42003-025-09440-7

Figure Lengend Snippet: A Representative images of H&E staining in tumor and adjacent normal tissues from FFPE specimens of three surgical SCLC patients. B IHC staining of RASA4 in tumor and adjacent normal tissues from FFPE specimens of the same three SCLC patients. The results show significantly reduced RASA4 expression in the tumor tissues compared to the adjacent normal tissues. C IHC staining of SERPINE2 in tumor and adjacent normal tissues from FFPE specimens of the same three SCLC patient samples. The results show significantly increased SERPINE2 expression in the tumor tissues compared to the adjacent normal tissues. (scale bar = 20 μm for 40×, scale bar = 100 μm for 8×) ( D ) IHC staining shows that SERPINE2 expression is elevated in the invasive front of the SCLC tumor. (scale bar = 40 μm for 20×, scale bar = 200 μm for 5×) ( E ) IHC scores of RASA4 and SERPINE2 in tumor and adjacent normal tissues from 29 SCLC patients who underwent surgery ( n = 29, Mann–Whitney test). F Statistical analysis indicates the negative association between RASA4 and SERPINE2 expression in SCLC tumor and adjacent normal tissues ( n = 29, Chi-square test for R × C contingency tables). G Kaplan–Meier survival analysis of 29 SCLC patients who underwent surgery, stratified by RASA4 IHC scores in tumor tissues. The results show that SCLC patients with moderate scores of RASA4 expression have significantly better survival compared to those with weak scores. The statistical significance of the difference in survival was determined by the log-rank test.

Article Snippet: SERPINE2 concentration in cell culture medium was quantified using the Human Glia-Derived Nexin (SERPINE2) ELISA Kit (#CSB-EL021082HU, Cusabio, Wuhan, China) following the manufacturer’s protocol.

Techniques: Staining, Immunohistochemistry, Expressing, MANN-WHITNEY

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