Journal: Gene

Article Title: Study on the mechanism of S100A4-mediated cancer oncogenesis in uveal melanoma cells through the integration of bioinformatics and in vitro experiments.

doi: 10.1016/j.gene.2024.148333

Figure Lengend Snippet: Fig. 9. S100A4 silencing induces UM cell apoptosis. (a–b) Annexin V-FITC/PI double fluorescence staining of UM cells was used to detect the apoptosis rate. (c–d) Hoechst 33342 staining was used to observe the change in cell apoptosis. (e–f) The effects of S100A4 silencing on apoptosis-related proteins were detected using western blotting. Significant differences are indicated as follows: *P < 0.05; **P < 0.01.

Article Snippet: Proteins were then transferred to polyvinylidene difluoride membranes, blocked for 1 h, and incubated with the following primary antibodies at the indicated dilution ratios: S100A4 (1:1000, cat. ab197896, Abcam, UK), N-cadherin (1:1000, cat. 66219–1-Ig, Proteintech, USA), E-cadherin (1:1000, cat. 60335–1-Ig, Proteintech, USA), vimentin (1:1000, cat. 60330–1-Ig; Proteintech, USA), proliferating cell nuclear antigen (PCNA, 1:1000, cat. 60097–1-Ig, Proteintech, USA), X-linked inhibitor of apoptosis protein (XIAP, 1:2000, cat. 66800–1-Ig; Proteintech, USA), survivin (1:1000, cat. 66495–1-Ig, Proteintech, USA), Bax (1:1000, cat. 60267–1-Ig, Proteintech, USA), Bcl-2 (1:1000, cat. ab196495, Abcam, UK), caspase-3 (1:1000, cat. ab184787, Abcam, UK), and GAPDH (1:5000, cat. 60004–1-Ig, Proteintech, USA).

Techniques: Fluorescence, Staining, Western Blot

Journal: OncoTargets and therapy

Article Title: Piwi-Interacting RNA1037 Enhances Chemoresistance and Motility in Human Oral Squamous Cell Carcinoma Cells

doi: 10.2147/OTT.S233322

Figure Lengend Snippet: Targeting the upregulated expression of piR-1037 induced by CDDP enhanced the sensitivity of OSCC cells to CDDP. ( A ) Confirmation of the suppressive effect of piRNA-1037 antisense oligonucleotides (piR-1037 complementary DNA oligonucleotides) (500 nM) on the expression of piRNA-1037 in SCC4 and SCC9 cells receiving the indicated treatments. ( B ) Western blot analysis and quantification of the effect of piR-1037 antisense oligonucleotides on the expression of the chemoresistance biomarkers MDR1 and α-enolase in SCC4 and SCC9 cells treated with CDDP (10 μM). ( C ) Decreased cell viability and increased apoptosis (TUNEL assay) ( D ) in OSCC cells transfected with piR-1037 antisense oligonucleotides and treated with CDDP compared with CDDP-treated cells transfected with a scrambled control. Cell apoptosis was detected using the TUNEL-based TiterTACS™ Colorimetric Apoptosis Detection Kit (Trevigen, USA). ( E ) Activity of caspase-3, caspase-8 and caspase-1 ( F ). The activity of caspases measured using Caspase-3 and Caspase-1 assay kits (Abcam, USA) and a Caspase-8 colorimetric kit (Sigma Aldrich, USA). The data represent the mean ±SD from at least three independent replicates. Statistical analysis was performed using one-way ANOVA analysis or unpaired student’s t -test: * p <0.05; ** p < 0.01.

Article Snippet: The membrane was then incubated with 5% nonfat milk in TBST (10 mM Tris, pH 8.0, 150 mM NaCl, 0.5% Tween 20) for 1 h. The membrane was washed once with TBST, followed by incubation with primary antibodies against MDR1 (Novus Biologicals, USA) (1:1000), α-enolase (Boster, China) (1:1000), XIAP (Abcam, USA) (1 μg/mL), cleaved caspase-3 (R&D systems, USA) (0,5 μg/mL), E-Cadherin (1:500), N-Cadherin (1:1000) (Abcam, USA), and β-actin (1:10,000) (Sigma Aldrich, USA).

Techniques: Expressing, Western Blot, TUNEL Assay, Transfection, Control, Activity Assay

Journal: Nature Communications

Article Title: Macrophage lineage cells-derived migrasomes activate complement-dependent blood-brain barrier damage in cerebral amyloid angiopathy mouse model

doi: 10.1038/s41467-023-39693-x

Figure Lengend Snippet: a Plasma CD5L concentration of CAA patients ( N = 12) and HC ( N = 12) as well as CAA models ( N = 5 in each group) and WT C57/BL6 mice ( N = 5). Data are presented as mean values ± SEM with the indicated significance (by two-tailed Student’s t test). b Left: CD5L deposition in mice brain was evaluated with immunostaining. Right: Immunostaining of CD31 (red), CD5L (green), C5b-9 (blue) and TSPAN4 (purple). Data are representative of 4 biologically independent experiments. c Plasma C5-9 concentration of mice. N = 6 in PBS-M, Aβ40-M and CD5L KO Aβ40-M transferred mice and N = 5 in CD5L KO PBS-M treated group. Data are presented as mean values ± SEM with the indicated significance (by one-way ANOVA followed by Tukey’s post-test). d C5b-9 deposition in brain as assessed with immunostaining. Data are representative of 3 biologically independent experiments. e Fluorescent intensity of 3kDa-Dextran in the plasma and eyeballs of the recipients. N = 6 in PBS-M, Aβ40-M and CD5L KO Aβ40-M transferred mice and N = 5 in CD5L KO PBS-M treated group. Data are presented as mean values ± SEM with the indicated significance (by one-way ANOVA followed by Tukey’s post-test). f Leakage of 3kDa-Dextran in brain parenchyma was assessed with fluorescent microscopy. Data are representative of 3 biologically independent experiments. g Coronal brain sections of the recipients were subjected to immunostaining of CD31 (green) and ZO-1 (red). Data are representative of 3 biologically independent experiments. h Skin autopsy sample of a CAA patient (Left) and brain tissue of WT and Tg-SwDI/B +/+ mice (24w of age) (Right) were subjected to IEM with CD5L staining. Blue dash lines outline micro-vessel wall. Red stars emphasize blood vessel lumen. Red arrow heads emphasize peri-vessel macrophages. Red arrows emphasize CD5L deposition in blood vessels within migrasome-like structures. Data are representative of 3 biologically independent experiments. Source data are provided as a Source Data file.

Article Snippet: Concentration of mouse NEFL (FineTest, EM1688), mouse C5b-9 (FineTest, EM1392), mouse CD5L (Boster, EK1414), human CD5L (MEIMIAN, MM-50587H1), human Aβ40 (CUSABIO, CSB-E08299h), human Aβ42 (MAI Bio, LM-2685H), and human C5b-9 (FineTest, EH3858) in plasma was assessed with commercial kits according to instructions of manufacturers.

Techniques: Clinical Proteomics, Concentration Assay, Two Tailed Test, Immunostaining, Microscopy, Staining

Journal: Translational Andrology and Urology

Article Title: Identification of BIRC5 and HMMR as prognostic biomarkers for immune infiltration in prostate cancer

doi: 10.21037/tau-24-359

Figure Lengend Snippet: Association between the expression of BIRC5 and HMMR genes and cancer prognosis. (A) Venn diagram revealing the identification of 159 DEGs at the intersection among three subgroups of PCa. (B) KEGG pathway enrichment analysis results for the 159 DEGs. (C) Cytoscape MCODE module used to screen 10 hub-key genes. (D,E) Kaplan-Meier curves of BCRFS for BIRC5 and HMMR in TCGA-PRAD. (F,G) Kaplan-Meier curves of BCRFS for BIRC5 and HMMR in GSE70769. (H,I) Univariate Cox regression analysis for differentially expressed hub-key genes in TCGA-PRAD and GSE70769. PCa, prostate cancer; mPCa, metastatic prostate cancer; CSPC, castration-sensitive prostate cancer; CRPC, castration-resistant prostate cancer; NEPC, neuroendocrine prostate cancer; ECM, extracellular matrix; BCRFS, biochemical recurrence-free survival; HR, hazard ratio; TCGA, The Cancer Genome Atlas; PRAD, prostate adenocarcinoma; DEGs, differentially expressed genes; KEGG, Kyoto Encyclopedia of Genes and Genomes.

Article Snippet: Primary antibodies for BIRC5 (Survivin, dilution 1:100, A00379, Boster, Wuhan, China) and HMMR (dilution 1:100, PA12592, Boster, Wuhan, China) were applied and incubated overnight at 4 °C.

Techniques: Expressing

Journal: Translational Andrology and Urology

Article Title: Identification of BIRC5 and HMMR as prognostic biomarkers for immune infiltration in prostate cancer

doi: 10.21037/tau-24-359

Figure Lengend Snippet: Analysis of the expression levels of BIRC5 and HMMR. (A) BIRC5 expression in tumor and normal tissues from TCGA pan-cancer and GTEx data. (B) HMMR expression in tumor and normal tissues from TCGA pan-cancer and GTEx data. (C,D) BIRC5 and HMMR expression in mPCa, localized PCa, and benign tissues from GSE3325. (E,F) BIRC5 and HMMR expression in NEPC and CSPC/CRPC tissues from GSE59984. (G) BIRC5 expression in mCRPC and localized PCa tissues from GSE35988. (H) HMMR expression in mCRPC and localized PCa tissues from GSE32269. *, P<0.05; **, P<0.01; ***, P<0.001; ns, not significant. TPM, transcripts per million; PCa, prostate cancer; mPCa, metastatic prostate cancer; CSPC, castration-sensitive prostate cancer; CRPC, castration-resistant prostate cancer; NEPC, neuroendocrine prostate cancer; PC, principal component; mCRPC, metastatic castration-resistant prostate cancer; TCGA, The Cancer Genome Atlas; GTEx, Genotype-Tissue Expression.

Article Snippet: Primary antibodies for BIRC5 (Survivin, dilution 1:100, A00379, Boster, Wuhan, China) and HMMR (dilution 1:100, PA12592, Boster, Wuhan, China) were applied and incubated overnight at 4 °C.

Techniques: Expressing

Journal: Translational Andrology and Urology

Article Title: Identification of BIRC5 and HMMR as prognostic biomarkers for immune infiltration in prostate cancer

doi: 10.21037/tau-24-359

Figure Lengend Snippet: Correlation and enrichment analyses of BIRC5 and HMMR. (A,B) Correlation between BIRC5, HMMR, and various immune cells; (C-H) association between the expression levels of BIRC5, HMMR, and the enrichment scores of various immune cells. *, P<0.05; **, P<0.01; ***, P<0.001.

Article Snippet: Primary antibodies for BIRC5 (Survivin, dilution 1:100, A00379, Boster, Wuhan, China) and HMMR (dilution 1:100, PA12592, Boster, Wuhan, China) were applied and incubated overnight at 4 °C.

Techniques: Expressing

Journal: Translational Andrology and Urology

Article Title: Identification of BIRC5 and HMMR as prognostic biomarkers for immune infiltration in prostate cancer

doi: 10.21037/tau-24-359

Figure Lengend Snippet: Correlation analysis between BIRC5 and HMMR expression and T helper cell and CD4 + /CD8 + T cell infiltration. (A-C) Correlation between BIRC5 expression in distinct PCa subgroups and Th cell infiltration levels using the xCELL algorithm. (D-F) Correlation between HMMR expression in distinct PCa subgroups and Th cell infiltration levels using the xCELL algorithm. (G-I) Correlation between BIRC5 expression in distinct PCa subgroups and CD4 + /CD8 + T cell infiltration levels using the quanTIseq algorithm. (J-L) Correlation between HMMR expression in distinct PCa subgroups and CD4 + /CD8 + T cell infiltration levels using the quanTIseq algorithm. PCa, prostate cancer; mCRPC, metastatic castration-resistant PCa; NEPC, neuroendocrine prostate cancer.

Article Snippet: Primary antibodies for BIRC5 (Survivin, dilution 1:100, A00379, Boster, Wuhan, China) and HMMR (dilution 1:100, PA12592, Boster, Wuhan, China) were applied and incubated overnight at 4 °C.

Techniques: Expressing

Journal: Translational Andrology and Urology

Article Title: Identification of BIRC5 and HMMR as prognostic biomarkers for immune infiltration in prostate cancer

doi: 10.21037/tau-24-359

Figure Lengend Snippet: Protein expression of BIRC5 and HMMR in PCa. (A-F) IHC images demonstrating the staining intensity of BIRC5 and HMMR in normal prostate tissues, Gleason ≤7 group, and Gleason >7 group (400×). (G,H) The AOD of IHC images of BIRC5 and HMMR used for comparisons among normal prostate tissues, the Gleason ≤7 group, and the Gleason >7 group. *, P<0.05; **, P<0.01; ***, P<0.001; ****, P<0.0001; AOD, average optical density; IHC, immunohistochemistry.

Article Snippet: Primary antibodies for BIRC5 (Survivin, dilution 1:100, A00379, Boster, Wuhan, China) and HMMR (dilution 1:100, PA12592, Boster, Wuhan, China) were applied and incubated overnight at 4 °C.

Techniques: Expressing, Staining, Immunohistochemistry