Journal: Biomedicines

Article Title: Podocalyxin, Isthmin-1, and Pentraxin-3 Immunoreactivities as Emerging Immunohistochemical Markers of Fibrosis in Chronic Hepatitis B

doi: 10.3390/biomedicines13122958

Figure Lengend Snippet: The immunoreactivities of PODXL, PTX3 and ISM-1 in the low fibrosis, high fibrosis, and control groups. (Arrows indicate immunoreactivities).

Article Snippet: After washing the tissues three times for 5 min each with PBS, 5 min of Ultra V Block (TA–125-UB, Lab Vision Corporation) solution was applied to prevent background staining, followed by incubation with PTX3, PODXL, and ISM-1 primary antibodies diluted at a ratio of 1/200 (PTX3 Polyclonal Antibody, PA5-36156, Thermo Fisher Scientific, Invitrogen, Waltham, MA, USA/PODXL, 39-3800, Thermo Fisher Scientific Inc./ISM1 Polyclonal Antibody, E-AB-18133, Elabscience Biotechnology, Houston, TX, USA) for 60 min at room temperature in a humidified environment.

Techniques: Control

Journal: Journal of Neuroinflammation

Article Title: K284-6111 alleviates memory impairment and neuroinflammation in Tg2576 mice by inhibition of Chitinase-3-like 1 regulating ERK-dependent PTX3 pathway

doi: 10.1186/s12974-020-02022-w

Figure Lengend Snippet: Effect of K284-6111 on PTX3-involved neuroinflammation. a Gene network analysis associated with CHI3L1 was carried out using the web-based analysis tool. BV-2 cells were treated with Aβ (5 μM) and K284-6111 (2 μM). (B) The mRNA expression level of Chi3l1 and Ptx3 in BV-2 cells was assessed by qRT-PCR. Expression of PTX3 was detected by Western blot ( c ) in the Tg2576 mice brain and ( d ) in the BV-2 cells. BV-2 cells were transfected with PTX3 siRNA (20 nM). After 24 hr, cells were treated with Aβ (5 μM). e The mRNA expression level of pro-inflammatory cytokines ( Tnf , Il1b , and Il6 ) and M1 microglia phenotype marker ( Cd86 ) in BV-2 cells were assessed by qRT-PCR. f Expression of iNOS and COX-2 was detected by Western blot. (G) Level of p-ERK 1/2, ERK 1/2, p-IκBα, and IκBα were detected by Western blot. BV-2 cells were treated with Aβ (5 μM), K284-6111 (2 μM), U0126 (20 μM), and Bay 11-7082 (5 μM). (H) Expression of PTX3 were detected by Western blot. BV-2 cells were transfected with CHI3L1 plasmid vector. After 24 hr, cells were treated with Aβ (5 μM), K284-6111 (2 μM), U0126 (20 μM), and Bay 11-7082 (5 μM). (I) Expression of PTX3 were detected by Western blot.

Article Snippet: Negative control (NC), PTX3 siRNA were purchased from OriGene Technologies, Inc. (Rockville, MD, USA). pcDNA3.1(+)-6 × Myc-CHI3L1 vector was cloned from Bionics (Seoul, Republic of Korea).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Transfection, Marker, Plasmid Preparation

Journal: Journal of Neuroinflammation

Article Title: K284-6111 alleviates memory impairment and neuroinflammation in Tg2576 mice by inhibition of Chitinase-3-like 1 regulating ERK-dependent PTX3 pathway

doi: 10.1186/s12974-020-02022-w

Figure Lengend Snippet: CHI3L1 mediates neuroinflammation through NF-κB pathway and ERK dependent PTX3 pathway

Article Snippet: Negative control (NC), PTX3 siRNA were purchased from OriGene Technologies, Inc. (Rockville, MD, USA). pcDNA3.1(+)-6 × Myc-CHI3L1 vector was cloned from Bionics (Seoul, Republic of Korea).

Techniques:

Journal: British journal of cancer

Article Title: Clinical impact of pentraxin family expression on prognosis of pancreatic carcinoma.

doi: 10.1038/bjc.2013.348

Figure Lengend Snippet: Figure 1. (A) Pentraxin 3 production levels of supernatants used for culturing of pancreatic carcinoma cell lines. (B, C) Cell migration assay in the presence or absence of the indicated reagents. NT (non-treated), FBS, 20% in the lower chamber; VEGF, 50 ng ml 1 in the lower chamber; PTX3, 50–100 ng ml 1; CRP, 50–100 ng ml 1. Statistical significance was evaluated by comparison with or without the presence of PTX3 and CRP.

Article Snippet: We used the transfectant pCMV6-entry PTX3 open reading frame (ORF) clones (OriGene Technologies, Inc., Rockville, MD, USA; cat. no. RC207922) to investigate the cellular activity induced by intracellular PTX3 overexpression.

Techniques: Cell Migration Assay, Comparison

Journal: British journal of cancer

Article Title: Clinical impact of pentraxin family expression on prognosis of pancreatic carcinoma.

doi: 10.1038/bjc.2013.348

Figure Lengend Snippet: Figure 3. Kaplan–Meier curves for (A) progression-free survival according to blood-PTX3 level and (B) overall survival according to blood-PTX3 level. Cutoff points for PTX3 level were based on mean PTX3 level.

Article Snippet: We used the transfectant pCMV6-entry PTX3 open reading frame (ORF) clones (OriGene Technologies, Inc., Rockville, MD, USA; cat. no. RC207922) to investigate the cellular activity induced by intracellular PTX3 overexpression.

Techniques:

Journal: Acta Pharmaceutica Sinica. B

Article Title: Sphingosine kinase 1 promotes growth of glioblastoma by increasing inflammation mediated by the NF- κ B /IL-6/STAT3 and JNK/PTX3 pathways

doi: 10.1016/j.apsb.2022.09.012

Figure Lengend Snippet: PTX3 is regulated by SPHK1 and promotes inflammation and proliferation of GBM cells. (A) Heatmap of top-ranked DEGs after SPHK1 overexpression in U251 cells. (B) The mRNA and protein expression of PTX3 was increased in U251 cells after overexpression of SPHK1. (C) The mRNA and protein expression of PTX3 was decreased in A172 and U87-MG cells after knockdown of SPHK1. (D) The concentration of PTX3 was increased in U251 cells after overexpression of SPHK1. (E) Confocal immunofluorescence staining of SPHK1 and PTX3 showed that protein expression of PTX3 was increased after overexpression of SPHK1. (F) Data from GEPIA website ( http://gepia.cancer-pku.cn/ ) shows that expression of PTX3 is higher in GBM tissues than in healthy tissues. (G) Data from the Broad Institute website ( http://gdac.broadinstitute.org/ .) shows that the expression of PTX3 is higher in GBM tissues than in healthy tissues. Data from (H, I) Oncomine ( www.oncomine.org ) and (J) UALCAN ( http://ualcan.path.uab.edu/index.html ) website shows that the expression of PTX3 is higher in GBM tissues than in normal tissues. (K) Data from Betastasis ( https://www.betastasis.com/ ) and GEPIA ( http://gepia.cancer-pku.cn/ ) website shows that the overall survival rate in patients with high expression of PTX3 is lower than in those with low expression of PTX3. (L) Immunohistochemical results indicate that the expression of PTX3 was higher in GBM tissues than in normal tissues. Representative images showing the expression of PTX3 in GBM and adjacent normal tissues. The data are presented as mean ± SD, and the experiments were performed in triplicate.

Article Snippet: The human full-length SPHK1 or PTX3 cDNA clone (Myc-DDK-tagged) was purchased from OriGene Technologies (Rockville, USA).

Techniques: Over Expression, Expressing, Knockdown, Concentration Assay, Immunofluorescence, Staining, Immunohistochemical staining

Journal: Acta Pharmaceutica Sinica. B

Article Title: Sphingosine kinase 1 promotes growth of glioblastoma by increasing inflammation mediated by the NF- κ B /IL-6/STAT3 and JNK/PTX3 pathways

doi: 10.1016/j.apsb.2022.09.012

Figure Lengend Snippet: PTX3 mediates the function of SPHK1 in GBM cells. (A) Protein expression of PTX3 in normal vs . GBM cells. (B) Overexpression of PTX3 increases the ratio of p-STAT3/STAT3 and the release of IL-6, and promotes colony formation of U251-MG cells. (C) Knockdown of PTX3 decreases the ratio of p-STAT3/STAT3 and the release of IL-6, and inhibits colony formation of U138-MG cells. (D) Strategy for investigating PTX3 function after overexpression of SPHK1. (E) Knockdown of PTX3 in U251 cells with overexpressed SPHK1. (F) ELISA results show that silencing PTX3 impairs release of IL-6 caused by overexpression of SPHK1 in U251-MG cells. (G) Silencing PTX3 decreases colony formation caused by overexpression of SPHK1 in U251-MG cells. (H) Silencing PTX3 inhibits growth of U251 cells in 3D Matrigel caused by overexpression of SPHK1. EdU analysis (I) and CCK-8 analysis (J) show that silencing PTX3 inhibits proliferation of U251 cells caused by overexpression of SPHK1. (K) Results from confocal immunofluorescence show that SPHK1 and PTX3 were co-localized in the cytoplasm and nuclei in U251-MG and U87-MG cells. The data are presented as mean ± SD, and the experiments were performed in triplicate.

Article Snippet: The human full-length SPHK1 or PTX3 cDNA clone (Myc-DDK-tagged) was purchased from OriGene Technologies (Rockville, USA).

Techniques: Expressing, Over Expression, Knockdown, Enzyme-linked Immunosorbent Assay, CCK-8 Assay, Immunofluorescence

Journal: Acta Pharmaceutica Sinica. B

Article Title: Sphingosine kinase 1 promotes growth of glioblastoma by increasing inflammation mediated by the NF- κ B /IL-6/STAT3 and JNK/PTX3 pathways

doi: 10.1016/j.apsb.2022.09.012

Figure Lengend Snippet: SPHK1 promotes the expression of PTX3 through ATF3. (A) Strategy for identifying ATF3 as the key transcription factor in regulating the expression of PTX3. (B) In silico analysis of TCGA data shows that SPHK1 is highly correlated with PTX3. (C) Results from confocal immunofluorescence show that SPHK1 and ATF3 or ATF3 and PTX3 are co-localized in the nucleus in U251-MG and U87-MG cells. (D) Prediction of the binding site of ATF3 in the promoter region of PTX3 using PROMO website ( http://alggen.lsi.upc.es/cgi-bin/promo_v3/promo/promoinit.cgi?dirDB=TF_8.3 .). (E) ChIP-qPCR analysis shows that ATF3 binds to PTX3 promoter in U251 cells. (F) Gel electrophoresis of ChIP-qPCR products. (G) Confocal immunofluorescence assay revealing that overexpression of ATF3 increases PTX3 expression in U251-MG cells. (H) Western blot results show that silencing ATF3 impairs the effects of SPHK1 on the expression of ATF3, PTX3, and the ratio of p-STAT3/STAT3 in U251-MG cells. Silencing ATF3 impairs the effect of SPHK1 on the expression of PTX3 (I), colony formation (J), 3D growth (K) in U251-MG cells. The data are presented as mean ± SD, and the experiments were performed in triplicate.

Article Snippet: The human full-length SPHK1 or PTX3 cDNA clone (Myc-DDK-tagged) was purchased from OriGene Technologies (Rockville, USA).

Techniques: Expressing, In Silico, Immunofluorescence, Binding Assay, ChIP-qPCR, Nucleic Acid Electrophoresis, Over Expression, Western Blot

Journal: Acta Pharmaceutica Sinica. B

Article Title: Sphingosine kinase 1 promotes growth of glioblastoma by increasing inflammation mediated by the NF- κ B /IL-6/STAT3 and JNK/PTX3 pathways

doi: 10.1016/j.apsb.2022.09.012

Figure Lengend Snippet: SPHK1 activates ATF3 through the JNK pathway and directly interacts with PTX3. (A) Strategy for identifying which pathway mediates the effects of SPHK1 on the regulation of PTX3. (B) Western blot results show that overexpression of SPHK1 increases the ratio of p-ERK/ERK and p-JNK/JNK in U251 cells. (C) Western blot results show that knockdown of SPHK1 and A172 cells reduces the ratio of P-ERK/ERK, p-JNK/JNK, and p-JUN/JUN in A172 and U87 cells. (D) The JNK inhibitor, SP600125, but not the ERK inhibitor, PD98059, suppresses expression of p-STAT3/STAT3, ATF3, and PTX3 after overexpression of SPHK1 in U251-MG cells. The JNK inhibitor, SP600125, but not the ERK inhibitor, PD98059, suppresses the release of IL-6 (E), colony formation (F) and growth in 3D Matrigel (G) in U251-MG cells after overexpression of SPHK1. (H) Co-IP analysis shows that SPHK1 and PTX3 co-immunoprecipitated in U251-MG cells and GBM tissues. (I) Confocal immunofluorescence results show that SPHK1 and PTX3 co-localize in the cytoplasm and nuclei in U251-MG cells. (J) Overexpression of PTX3 increases the expression of ATF3, p-JUN/JUN, and p-JNK/JNK in U251 cells. (K) Overexpression of SPHK1 or PTX3 increases the expression of ATF3, p-JUN/JUN, and p-STAT3/STAT3 in the nuclei of U251 cells. The data are presented as mean ± SD, and the experiments were performed in triplicate.

Article Snippet: The human full-length SPHK1 or PTX3 cDNA clone (Myc-DDK-tagged) was purchased from OriGene Technologies (Rockville, USA).

Techniques: Western Blot, Over Expression, Knockdown, Expressing, Co-Immunoprecipitation Assay, Immunoprecipitation, Immunofluorescence

Journal: Acta Pharmaceutica Sinica. B

Article Title: Sphingosine kinase 1 promotes growth of glioblastoma by increasing inflammation mediated by the NF- κ B /IL-6/STAT3 and JNK/PTX3 pathways

doi: 10.1016/j.apsb.2022.09.012

Figure Lengend Snippet: SPHK1 promotes tumor growth and SPHK1 inhibitor, PF543, suppresses tumorigenesis in vivo . (A) Strategy for studying the tumor promotion effects of SPHK1 in vivo . (B) Overexpression of SPHK1 in U251-MG cells increases tumor volume and weight in nude mice. The data are presented as mean ± SD, n = 6 in each group. (C) Overexpression of SPHK1 in U251-MG cells increases the expression of PTX3 and S1P levels in nude mice. The data are presented as mean ± SD, n = 3 in each group. (D) Knockdown of SPHK1 in U87-MG cells suppresses tumor growth in nude mice. The data are presented as mean ± SD, n = 6 in each group. (E) Knockdown of SPHK1 in U87-MG cells decreases the expression of PTX3 and S1P levels in nude mice. The data are presented as mean ± SD, n = 3 in each group. (F) PF543 significantly reduces tumor volume in mice transplanted with U87-MG cells in the orthotopic model. The data are presented as mean ± SD, n = 3 for MRI and tumor volume and n = 6 for body weight. (G) PF543 significantly reduces the expression of PTX3 and S1P levels in mice transplanted with U87-MG cells in the orthotopic model. The data are presented as mean ± SD, n = 3 in each group. (H) PF543 significantly inhibits tumor growth in mice transplanted with U251-MG SPHK1 cells in the orthotopic model. The data are presented as mean ± SD, n = 3 in each group. (I) PF543 significantly reduces the expression of PTX3 and S1P levels in mice transplanted with U251-MG SPHK1 cells in the orthotopic model. The data are presented as mean ± SD, n = 3 for MRI and tumor volume and n = 6 for body weight.

Article Snippet: The human full-length SPHK1 or PTX3 cDNA clone (Myc-DDK-tagged) was purchased from OriGene Technologies (Rockville, USA).

Techniques: In Vivo, Over Expression, Expressing, Knockdown

Journal: Acta Pharmaceutica Sinica. B

Article Title: Sphingosine kinase 1 promotes growth of glioblastoma by increasing inflammation mediated by the NF- κ B /IL-6/STAT3 and JNK/PTX3 pathways

doi: 10.1016/j.apsb.2022.09.012

Figure Lengend Snippet: Diagram showing the proposed mechanism of SPHK1 action in GBM cells. SPHK1 promotes growth of GBM by increasing inflammation mediated by the NF- κ B/IL-6/STAT3 and JNK/PTX3 pathways.

Article Snippet: The human full-length SPHK1 or PTX3 cDNA clone (Myc-DDK-tagged) was purchased from OriGene Technologies (Rockville, USA).

Techniques: