Journal: Cell Death & Disease

Article Title: Inhibition of the immunoproteasome modulates innate immunity to ameliorate muscle pathology of dysferlin-deficient BlAJ mice

doi: 10.1038/s41419-022-05416-1

Figure Lengend Snippet: Proteomic analysis of IP subunits ( A ); alarmins and inflammatory mediators (PTX3 and GPx1) ( B ); MAPK kinases and AKT1/2/3 and their phosphorylated isoforms ( C ) in psoas of 6 m BlAJ, 12 m BlAJ and BlAJ+ONX, 12 m C57Bl mice. Psoas stained with isolectin (in green), PTX3 (in magenta) with nuclei stained in DAPI (blu) (scale bar: 10 μm) and its magnification (scale bar: 10 μm) with white arrows indicating cells co-expressing PTX3 and isolectin. The histogram represents the counting of PTX3 fluorescence ( D ). Immunohistological staining of skeletal muscles for Iba1 (in red) and CD206 (in green). Nuclei were counterstained with DAPI (blue). Scale bar: 50 μm. Quantification of macrophage percentages. Mac1 were stained with Iba1+ and Mac2 with CD206+ in 6 m BIAJ, 12 m BlAJ and BlAJ+ONX, 12 m C57Bl skeletal muscles ( E ). Data are presented as mean ± SD of n = 3 independent experiments with n = 3–6 animals/group. One-way ANOVA, Tukey multiple comparison test for WB and non-parametric test followed by Kruskal–Wallis test for PTX3+ cells’ and macrophages’ counting: * p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001.

Article Snippet: Samples were resolved on polyacrylamide gels (ranging from 6 to 12%), transferred to nitrocellulose membranes (Bio-Rad Laboratories, California, USA) and overnight incubated with following antibodies: vinculin (1:600, MA5-11690, Invitrogen); PTX3 (C-10 1:600, sc-373951, Santa Cruz Biotechnology); p38 (1:500, E-AB-32460, Elabscience); PSMB5 (1:500; ab3330, Abcam); PSMB8 (1:500, Proteasome 20S LMP7, ab3329, Abcam); PSMB9 (1:500; Proteasome 20S LMP2 (EPR13785) ab184172, Abcam); S100β chain (C-3) (1:500, sc-393919, Santa Cruz Biotechnology); TNFα (1:500, E-AB-40015, Elabscience); HMGB1 (HAP46.5) (1:600, sc-56698, Santa Cruz); GAPDH (0411) (1:600, sc-47724, Santa Cruz); TLR4 (1:500, sc-293072, Santa Cruz); phosphoP38 (1:500, E-AB-20949, Elabscience); ERK1-2 (1:500, ab54230, Abcam); phosphoERK1-2 (1:500, E-AB-20868, Elabscience); AMPK-1 α (1:500, sc-74461, Santa Cruz); phosphoAMPK-α1/2 (1:600, STJ-90735, STJohnlabs); GSK-3 αβ (1ː500, sc-81496, Santa Cruz); TLR2 (1500, orb229137, Biorbyt): mTOR (1:500, PA5-34663, Invitrogen); PKCα (1:600, 610108, BD); CDK4 (1:500, sc-23896, Santa Cruz); Cyclin D1 (1:500, sc-8396, Santa Cruz); Cyclin E (1:500, sc-377100, Santa Cruz); MMP2 (1:500, ab37150, Abcam); MMP9 (1:500, ab38898, Abcam); adiponectin (1:500, 710179, Invitrogen); ATG-7 (1:500, sab4200304, Sigma-Aldrich); LC3B (1:500, L7543, Sigma-Aldrich); DRP1 (1:500, AB184247, Abcam); OXPHOS (1:500, MS604-300, Abcam); PGC1α (1:500, sc-518038, Santa Cruz); P62 (1:500, P0067, Sigma-Aldrich); RAGE (1:500, NBP2-03950, Novusbio); MYD88 (1:500, 23230-1-AP, Proteintech); AKT1/2/3 (1:500, ab126811, Abcam); Perilipin-1 (1:600, ab3526, Abcam); C3aR1 (1:100, PA5-109467 Invitrogen); pAkt (Ser473)(1:600, sc-81515, Santa Cruz Biotechnology); IKK-I (1:600, sc-376114, Santa Cruz Biotechnology); C5aR1/CD88 (1:600, 21316-1-AP, Proteintech); GPx1 (1:600, ab22604, Abcam).

Techniques: Staining, Expressing, Fluorescence, Muscles, Comparison

Journal: CNS Neuroscience & Therapeutics

Article Title: Targeting blood brain barrier—Remote ischemic conditioning alleviates cognitive impairment in female APP / PS1 rats

doi: 10.1111/cns.14613

Figure Lengend Snippet: Antibody information.

Article Snippet: PTX3 , 1:100 , 1:500 , Abclonal , R , AB_2759517 , A12670.

Techniques:

Journal: CNS Neuroscience & Therapeutics

Article Title: Targeting blood brain barrier—Remote ischemic conditioning alleviates cognitive impairment in female APP / PS1 rats

doi: 10.1111/cns.14613

Figure Lengend Snippet: Primers for qPCR.

Article Snippet: PTX3 , 1:100 , 1:500 , Abclonal , R , AB_2759517 , A12670.

Techniques:

Journal: CNS Neuroscience & Therapeutics

Article Title: Targeting blood brain barrier—Remote ischemic conditioning alleviates cognitive impairment in female APP / PS1 rats

doi: 10.1111/cns.14613

Figure Lengend Snippet: Remote ischemia conditioning reversed A1‐A2 astrocyte polarization in the frontal–parietal cortex of OVX APP/PS1 rats. (A‐a1–a5) Astrocyte activation was confirmed by examining the protein expression of PAN marker vimentin, and two typical astrocyte markers, C3 (A1), S100A10, PTX3, and Stat3 (A2) using Western blot analysis. (a6) Double immunofluorescence staining showed increased co‐localization of PTX3 with GFAP in RIC‐treated rats compared to the APP/PS1 group. (B–D) RT‐qPCR analysis showed gene levels of vimentin and A1/A2 markers of astrocytes. Values are expressed as means ± SEM of determinations from each group. N = 4–6, magnification 40×, scale bar 50 μm.

Article Snippet: PTX3 , 1:100 , 1:500 , Abclonal , R , AB_2759517 , A12670.

Techniques: Activation Assay, Expressing, Marker, Western Blot, Double Immunofluorescence Staining, Quantitative RT-PCR

Journal: International Journal of Nanomedicine

Article Title: Proof of Concept of the Radiosensitizing Effect of Gadolinium Oxide Nanoparticles in Cell Spheroids and a Tumor-Implanted Murine Model of Chondrosarcoma

doi: 10.2147/IJN.S390056

Figure Lengend Snippet: Gene expression of PTX3, CXCR4, MYO5C or PLS1 and PTX3 protein expression in tumors of mice sacrificed 7 days after treatment. ( A ) Box plots of RNA expression counts (median ± 1.5 times the interquartile range) for PTX3, CXCR4, MYO5C or PLS1 genes in three tumors of each group of treatment (NaCl, NaCl + 4×1 Gy and GdoNP (IV) + 4×1 Gy). ( B ) Fluorescent immunostaining of PTX3 protein (red) and nuclei (DAPI) in tumor sections for one mouse from each group of treatments. (* p < 0.05, **p<0.01, ***p<0.005).

Article Snippet: Tumor slides were incubated with a polyclonal anti-PTX3 (#HPA069320 Atlas Antibodies Ozyme, St Quentin, France) and then with an Alexafluor 555-labelled donkey anti-rabbit (#ab150074 Abcam).

Techniques: Expressing, RNA Expression, Immunostaining

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: 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: CNS Neuroscience & Therapeutics

Article Title: Biphasic roles of pentraxin 3 in cerebrovascular function after white matter stroke

doi: 10.1111/cns.13510

Figure Lengend Snippet: Pentraxin 3 expression after white matter stroke: A‐B, Postmortem brain tissue with ischemic stroke showed upregulation of PTX3 in the peri‐infarct area of white matter, whereas tissues without stroke expressed infrequent PTX3 in subcortical white matter. Scale bar = 100 μm. Data are mean ± SD from N = 5. * P < .05 (unpaired t test with Welch's correction). A, C, Immunostaining showed that PTX3 expression was increased at day 3 after white matter stroke caused by ET‐1 injection in mice. Scale bar = 100 μm. D, Western blots showed that PTX3 increased after ET‐1 injection was sustained for up to at least 21 d. Data are mean ± SD from N = 5. * P < .05 vs sham (one‐way ANOVA followed by post hoc Dunnett's multiple comparison test). E, Double staining of PTX3 with cell‐specific marker antibodies showed that the major cell type of PTX3 expression at day 3 after ET‐1 injection was GFAP‐positive astrocytes. Scale bar = 50 μm

Article Snippet: N5‐(1‐iminoethyl)‐L‐ornithine (L‐Nio), control siRNA, and mouse PTX3 siRNA were purchased from Santa Cruz.

Techniques: Expressing, Immunostaining, Injection, Western Blot, Comparison, Double Staining, Marker

Journal: CNS Neuroscience & Therapeutics

Article Title: Biphasic roles of pentraxin 3 in cerebrovascular function after white matter stroke

doi: 10.1111/cns.13510

Figure Lengend Snippet: Pentraxin 3 and myelin density during the acute phase of white matter stroke in mice: Male mice were subjected to ET‐1 injection after the treatment of control siRNA or PTX3 siRNA. Three days after ET‐1 injection, white matter stroke mice were sacrificed, and brain samples were used for Western blotting or immunostaining. A, Western blotting confirmed that PTX3 siRNA downregulated PTX3 expression in the affected region at day 3 after ET‐1 injection. Data are mean ± SD from N = 4. * P < .05 (unpaired t test with Welch's correction). B, Myelin staining showed that ET‐1 injection caused myelin damage in the corpus callosum at day 3 after ET‐1 injection, and mice that received PTX3 siRNA exhibited less myelin density compared to mice with control siRNA. Scale bar = 100 μm. Data are mean ± SD from N = 8 for sham and N = 12 for ET‐1/control siRNA and ET‐1/PTX3‐siRNA. * P < .05 (one‐way ANOVA followed by post hoc Tukey's multiple comparison test)

Article Snippet: N5‐(1‐iminoethyl)‐L‐ornithine (L‐Nio), control siRNA, and mouse PTX3 siRNA were purchased from Santa Cruz.

Techniques: Injection, Control, Western Blot, Immunostaining, Expressing, Staining, Comparison

Journal: CNS Neuroscience & Therapeutics

Article Title: Biphasic roles of pentraxin 3 in cerebrovascular function after white matter stroke

doi: 10.1111/cns.13510

Figure Lengend Snippet: Pentraxin and BBB damage during the acute phase of white matter stroke in mice: Male mice were subjected to ET‐1 injection after the treatment of control siRNA or PTX3 siRNA. Three days after ET‐1 injection, white matter stroke mice were sacrificed, and brain samples were used for immunostaining. A, IgG staining showed that ET‐1 injection caused BBB damage in the corpus callosum at day 3 after ET‐1 injection, and mice that received PTX3 siRNA exhibited a larger extent of BBB damage compared to mice with control siRNA. Scale bar = 50 μm. Data are mean ± SD from N = 8 for sham and N = 12 for ET‐1/control siRNA and ET‐1/PTX3‐siRNA. * P < .05 (Kruskal‐Wallis followed by post hoc Steel‐Dwass test). B, Mice that received PTX3 siRNA showed more leakage of 70 kDa FITC dextran at day 3. Scale bar = 50 μm. Data are mean ± SD from N = 8 for sham and N = 12 for ET‐1/control siRNA and ET‐1/PTX3‐siRNA. * P < .05 (Kruskal‐Wallis followed by post hoc Steel‐Dwass test). C, Similarly, mice that received PTX3 siRNA also showed more leakage of TRITC albumin at day 3. Scale bar = 50 μm. Data are mean ± SD from N = 8 for sham and N = 12 for ET‐1/control siRNA and ET‐1/PTX3‐siRNA. * P < .05 (Kruskal‐Wallis followed by post hoc Steel‐Dwass test). D, Concomitantly, the expression level of a tight junction accessory protein ZO‐1 was lower in the mice with PTX3‐siRNA. Scale bar = 50 μm. Data are mean ± SD from N = 8 for sham and N = 12 for ET‐1/control siRNA and ET‐1/PTX3‐siRNA. Please see Figure for the confirmation that ZO‐1 expression was observed in CD31‐positive cells in mouse corpus callosum. * P < .05 (Kruskal‐Wallis followed by post hoc Steel‐Dwass test)

Article Snippet: N5‐(1‐iminoethyl)‐L‐ornithine (L‐Nio), control siRNA, and mouse PTX3 siRNA were purchased from Santa Cruz.

Techniques: Injection, Control, Immunostaining, Staining, Expressing

Journal: CNS Neuroscience & Therapeutics

Article Title: Biphasic roles of pentraxin 3 in cerebrovascular function after white matter stroke

doi: 10.1111/cns.13510

Figure Lengend Snippet: Pentraxin and compensatory angiogenesis in chronic phase of white matter stroke in mice: Male mice were subjected to ET‐1 injection, and 7 d later, these mice were treated with either control siRNA or PTX3 siRNA. As a negative control, mice were subjected to ET‐1 injection after the treatment of PTX3 siRNA. Twenty‐one days after ET‐1 injection, white matter stroke mice were sacrificed, and brain samples were used for Western blotting or immunostaining. Please see Figure for the experimental diagram of this experiment. A, Western blotting confirmed that when PTX3 siRNA was treated at day 7 after ET‐1 injection, PTX3 expression in the affected region was downregulated at day 10 after ET‐1 injection. Data are mean ± SD from N = 4. * P < .05 (unpaired t test with Welch's correction). B, CD31/BrdU‐double staining showed that mice with PTX3‐siRNA (@day 7) had a larger number of CD31/BrdU‐double positive cells (newly generated endothelial cells) at day 21 after white matter stroke, compared to mice with control siRNA (@day 7) or mice with PTX3‐siRNA (@day 0). The inset is an enlarged CD31/BrdU‐double staining image, which confirmed that some of BrdU‐positive cells were merged with CD31 staining. Scale bar = 100 μm. Data are mean ± SD from N = 12. * P < .05 vs Sham, # P < .05 vs ET‐1/Control siRNA (Kruskal‐Wallis followed by post hoc Steel‐Dwass test). C, Similarly, PTX3‐siRNA (@day 7) promoted compensatory angiogenesis at day 21 after ET‐1 injection. Scale bar = 100 μm. Data are mean ± SD from N = 8 for Sham and N = 16 for other groups. * P < .05 (Kruskal‐Wallis followed by post hoc Steel‐Dwass test)

Article Snippet: N5‐(1‐iminoethyl)‐L‐ornithine (L‐Nio), control siRNA, and mouse PTX3 siRNA were purchased from Santa Cruz.

Techniques: Injection, Control, Negative Control, Western Blot, Immunostaining, Expressing, Double Staining, Generated, Staining

Journal: CNS Neuroscience & Therapeutics

Article Title: Biphasic roles of pentraxin 3 in cerebrovascular function after white matter stroke

doi: 10.1111/cns.13510

Figure Lengend Snippet: Pentraxin and in vitro angiogenesis: Brain endothelial RBE.4 cells were used for the Matrigel tube formation assay. A, RBE.4 cells were maintained in culture medium containing recombinant PTX3 for 18 h. PTX3 treatment suppressed the tube formation in a dose‐response manner. Data are mean ± SD from N = 5. * P < .05 (one‐way ANOVA followed by post hoc Tukey's multiple comparison test). B, Recombinant VEGF (10 ng/mL for 18 h) promoted the tube formation, but cotreatment with recombinant PTX3 (10 ng/mL) suppressed VEGF‐induced tube formation. Data are mean ± SD from N = 5. * P < .05 (one‐way ANOVA followed by post hoc Tukey's multiple comparison test)

Article Snippet: N5‐(1‐iminoethyl)‐L‐ornithine (L‐Nio), control siRNA, and mouse PTX3 siRNA were purchased from Santa Cruz.

Techniques: In Vitro, Tube Formation Assay, Recombinant, Comparison