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rabbit polyclonal anti nlrp3  (Novus Biologicals)


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    Structured Review

    Novus Biologicals rabbit polyclonal anti nlrp3
    TCP increases SESN2 levels in NE mice cochleae. (A) Representative images of the medial cochlear turn in the control, NE+saline, and NE+TCP groups ( n = 3 cochleae from three mice per group). Hair cells were visualized by immunofluorescent staining for phalloidin (red), and the tissues were also stained for SESN2 (green), and nuclei (DAPI; blue). (B) Hair cells were visualized by immunofluorescent staining for phalloidin (red), and the tissues were also stained for <t>NLRP3</t> (green), and nuclei (DAPI; blue) (n=3 cochleae from three mice per group). Scale bars: 20 μm in A and B. (C) Quantification of SESN2- and NLRP3-positive cells among the three groups. (D) SESN2 expression in mice cochleae ( n = 3 cochleae from three mice per group). All experiments were replicated thrice. Data are presented as mean ± standard of the mean. * P < 0.05, ** P < 0.01, **** P < 0.0001 (one-way analysis of variance with Bonferroni’s multiple comparison test). Ctrl: Control; DAPI: 4,6-diamidino-2-phenylindole; NE: noise exposure; ns: not significant; TCP: tranylcypromine.
    Rabbit Polyclonal Anti Nlrp3, supplied by Novus Biologicals, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "Tranylcypromine upregulates Sestrin 2 expression to ameliorate NLRP3-related noise-induced hearing loss"

    Article Title: Tranylcypromine upregulates Sestrin 2 expression to ameliorate NLRP3-related noise-induced hearing loss

    Journal: Neural Regeneration Research

    doi: 10.4103/NRR.NRR-D-24-00130

    TCP increases SESN2 levels in NE mice cochleae. (A) Representative images of the medial cochlear turn in the control, NE+saline, and NE+TCP groups ( n = 3 cochleae from three mice per group). Hair cells were visualized by immunofluorescent staining for phalloidin (red), and the tissues were also stained for SESN2 (green), and nuclei (DAPI; blue). (B) Hair cells were visualized by immunofluorescent staining for phalloidin (red), and the tissues were also stained for NLRP3 (green), and nuclei (DAPI; blue) (n=3 cochleae from three mice per group). Scale bars: 20 μm in A and B. (C) Quantification of SESN2- and NLRP3-positive cells among the three groups. (D) SESN2 expression in mice cochleae ( n = 3 cochleae from three mice per group). All experiments were replicated thrice. Data are presented as mean ± standard of the mean. * P < 0.05, ** P < 0.01, **** P < 0.0001 (one-way analysis of variance with Bonferroni’s multiple comparison test). Ctrl: Control; DAPI: 4,6-diamidino-2-phenylindole; NE: noise exposure; ns: not significant; TCP: tranylcypromine.
    Figure Legend Snippet: TCP increases SESN2 levels in NE mice cochleae. (A) Representative images of the medial cochlear turn in the control, NE+saline, and NE+TCP groups ( n = 3 cochleae from three mice per group). Hair cells were visualized by immunofluorescent staining for phalloidin (red), and the tissues were also stained for SESN2 (green), and nuclei (DAPI; blue). (B) Hair cells were visualized by immunofluorescent staining for phalloidin (red), and the tissues were also stained for NLRP3 (green), and nuclei (DAPI; blue) (n=3 cochleae from three mice per group). Scale bars: 20 μm in A and B. (C) Quantification of SESN2- and NLRP3-positive cells among the three groups. (D) SESN2 expression in mice cochleae ( n = 3 cochleae from three mice per group). All experiments were replicated thrice. Data are presented as mean ± standard of the mean. * P < 0.05, ** P < 0.01, **** P < 0.0001 (one-way analysis of variance with Bonferroni’s multiple comparison test). Ctrl: Control; DAPI: 4,6-diamidino-2-phenylindole; NE: noise exposure; ns: not significant; TCP: tranylcypromine.

    Techniques Used: Control, Saline, Staining, Expressing, Comparison



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    TCP increases SESN2 levels in NE mice cochleae. (A) Representative images of the medial cochlear turn in the control, NE+saline, and NE+TCP groups ( n = 3 cochleae from three mice per group). Hair cells were visualized by immunofluorescent staining for phalloidin (red), and the tissues were also stained for SESN2 (green), and nuclei (DAPI; blue). (B) Hair cells were visualized by immunofluorescent staining for phalloidin (red), and the tissues were also stained for <t>NLRP3</t> (green), and nuclei (DAPI; blue) (n=3 cochleae from three mice per group). Scale bars: 20 μm in A and B. (C) Quantification of SESN2- and NLRP3-positive cells among the three groups. (D) SESN2 expression in mice cochleae ( n = 3 cochleae from three mice per group). All experiments were replicated thrice. Data are presented as mean ± standard of the mean. * P < 0.05, ** P < 0.01, **** P < 0.0001 (one-way analysis of variance with Bonferroni’s multiple comparison test). Ctrl: Control; DAPI: 4,6-diamidino-2-phenylindole; NE: noise exposure; ns: not significant; TCP: tranylcypromine.
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    TCP increases SESN2 levels in NE mice cochleae. (A) Representative images of the medial cochlear turn in the control, NE+saline, and NE+TCP groups ( n = 3 cochleae from three mice per group). Hair cells were visualized by immunofluorescent staining for phalloidin (red), and the tissues were also stained for SESN2 (green), and nuclei (DAPI; blue). (B) Hair cells were visualized by immunofluorescent staining for phalloidin (red), and the tissues were also stained for <t>NLRP3</t> (green), and nuclei (DAPI; blue) (n=3 cochleae from three mice per group). Scale bars: 20 μm in A and B. (C) Quantification of SESN2- and NLRP3-positive cells among the three groups. (D) SESN2 expression in mice cochleae ( n = 3 cochleae from three mice per group). All experiments were replicated thrice. Data are presented as mean ± standard of the mean. * P < 0.05, ** P < 0.01, **** P < 0.0001 (one-way analysis of variance with Bonferroni’s multiple comparison test). Ctrl: Control; DAPI: 4,6-diamidino-2-phenylindole; NE: noise exposure; ns: not significant; TCP: tranylcypromine.
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    TCP increases SESN2 levels in NE mice cochleae. (A) Representative images of the medial cochlear turn in the control, NE+saline, and NE+TCP groups ( n = 3 cochleae from three mice per group). Hair cells were visualized by immunofluorescent staining for phalloidin (red), and the tissues were also stained for SESN2 (green), and nuclei (DAPI; blue). (B) Hair cells were visualized by immunofluorescent staining for phalloidin (red), and the tissues were also stained for <t>NLRP3</t> (green), and nuclei (DAPI; blue) (n=3 cochleae from three mice per group). Scale bars: 20 μm in A and B. (C) Quantification of SESN2- and NLRP3-positive cells among the three groups. (D) SESN2 expression in mice cochleae ( n = 3 cochleae from three mice per group). All experiments were replicated thrice. Data are presented as mean ± standard of the mean. * P < 0.05, ** P < 0.01, **** P < 0.0001 (one-way analysis of variance with Bonferroni’s multiple comparison test). Ctrl: Control; DAPI: 4,6-diamidino-2-phenylindole; NE: noise exposure; ns: not significant; TCP: tranylcypromine.
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    TCP increases SESN2 levels in NE mice cochleae. (A) Representative images of the medial cochlear turn in the control, NE+saline, and NE+TCP groups ( n = 3 cochleae from three mice per group). Hair cells were visualized by immunofluorescent staining for phalloidin (red), and the tissues were also stained for SESN2 (green), and nuclei (DAPI; blue). (B) Hair cells were visualized by immunofluorescent staining for phalloidin (red), and the tissues were also stained for <t>NLRP3</t> (green), and nuclei (DAPI; blue) (n=3 cochleae from three mice per group). Scale bars: 20 μm in A and B. (C) Quantification of SESN2- and NLRP3-positive cells among the three groups. (D) SESN2 expression in mice cochleae ( n = 3 cochleae from three mice per group). All experiments were replicated thrice. Data are presented as mean ± standard of the mean. * P < 0.05, ** P < 0.01, **** P < 0.0001 (one-way analysis of variance with Bonferroni’s multiple comparison test). Ctrl: Control; DAPI: 4,6-diamidino-2-phenylindole; NE: noise exposure; ns: not significant; TCP: tranylcypromine.
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    <t>NLRP3</t> expression in EOC and Kaplan-Meier survival analyses. ( a ) A box plot showed differential NLRP3 expression in normal and EOC samples. ( b ) Immunohistochemistry verified NLRP3 expression patterns in normal ovary (n=10) and EOC tissues (n=10) and their AODs; ( c ) Representative Western blot image showing NLRP3 protein expression in normal ovarian epithelial cells (IOSE80) and various EOC cell lines. Quantitative analysis of protein expression levels is shown below, with data represented as mean ± SD from three independent experiments. Symbols indicate statistically significant differences between IOSE80 and ovarian cancer cell lines. ( d – f ) Kaplan-Meier analysis revealed significant differences in patients’ DFS ( d ), PFS ( e ), and OS ( f ) between NLRP3 high and low expression groups in the training cohort. Data are shown as mean±SD; ns, no significance; ***, p < 0.001; ****, p < 0.0001.
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    <t>NLRP3</t> expression in EOC and Kaplan-Meier survival analyses. ( a ) A box plot showed differential NLRP3 expression in normal and EOC samples. ( b ) Immunohistochemistry verified NLRP3 expression patterns in normal ovary (n=10) and EOC tissues (n=10) and their AODs; ( c ) Representative Western blot image showing NLRP3 protein expression in normal ovarian epithelial cells (IOSE80) and various EOC cell lines. Quantitative analysis of protein expression levels is shown below, with data represented as mean ± SD from three independent experiments. Symbols indicate statistically significant differences between IOSE80 and ovarian cancer cell lines. ( d – f ) Kaplan-Meier analysis revealed significant differences in patients’ DFS ( d ), PFS ( e ), and OS ( f ) between NLRP3 high and low expression groups in the training cohort. Data are shown as mean±SD; ns, no significance; ***, p < 0.001; ****, p < 0.0001.
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    <t>NLRP3</t> expression in EOC and Kaplan-Meier survival analyses. ( a ) A box plot showed differential NLRP3 expression in normal and EOC samples. ( b ) Immunohistochemistry verified NLRP3 expression patterns in normal ovary (n=10) and EOC tissues (n=10) and their AODs; ( c ) Representative Western blot image showing NLRP3 protein expression in normal ovarian epithelial cells (IOSE80) and various EOC cell lines. Quantitative analysis of protein expression levels is shown below, with data represented as mean ± SD from three independent experiments. Symbols indicate statistically significant differences between IOSE80 and ovarian cancer cell lines. ( d – f ) Kaplan-Meier analysis revealed significant differences in patients’ DFS ( d ), PFS ( e ), and OS ( f ) between NLRP3 high and low expression groups in the training cohort. Data are shown as mean±SD; ns, no significance; ***, p < 0.001; ****, p < 0.0001.
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    Novus Biologicals rabbit polyclonal anti nlrp3 nalp3 antibody
    In vitro characterization of a DNA vector expressing the multiepitopic CoV2-TMEP protein. ( A ) Schematic representation of the chimeric CoV2-TMEP multiepitopic vaccine targeting T cells. ( B ) Time-course expression of CoV2-TMEP protein. HEK-293T cells were transfected as detailed in the Materials and Methods section and CoV2-TMEP expression in pellets and supernatants (SN) was analyzed by Western blotting using the mouse monoclonal anti-FLAG antibody. ( C ) Subcellular localization of CoV2-TMEP protein. HeLa cells were transfected and processed for immunofluorescence microscopy as indicated in the Materials and Methods section. Green staining: mouse monoclonal anti-FLAG antibody followed by anti-mouse Alexa Fluor 488 antibody. Yellow arrows: CoV2-TMEP forming aggregates in magnified images (10×). ( D ) Analysis of <t>NLRP3</t> induction and PARP cleavage. HEK-293T cells were transfected as described in the Materials and Methods section and the expression of NLRP3 (118 KDa; upper panel) and PARP (full-length: 116 KDa; cleaved: 89 KDa; lower panel) was analyzed by Western blotting using specific antibodies. ( E ) Subcellular colocalization of CoV2-TMEP protein with inflammasome. HeLa cells were transfected and processed for confocal microscopy as detailed in the Materials and Methods section. A mouse monoclonal anti-FLAG antibody and a rabbit polyclonal <t>anti-NLRP3/NALP3</t> antibody followed by the corresponding secondary antibodies conjugated with Alexa Fluor 488 (green staining; FLAG detection) or Alexa Fluor 594 (red staining; NLRP3 detection) were used. Colocalization signal is indicated in white. Yellow arrows: Discrete colocalization signals in magnified images (5×). One representative optical section from three independent experiments is shown.
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    Image Search Results


    TCP increases SESN2 levels in NE mice cochleae. (A) Representative images of the medial cochlear turn in the control, NE+saline, and NE+TCP groups ( n = 3 cochleae from three mice per group). Hair cells were visualized by immunofluorescent staining for phalloidin (red), and the tissues were also stained for SESN2 (green), and nuclei (DAPI; blue). (B) Hair cells were visualized by immunofluorescent staining for phalloidin (red), and the tissues were also stained for NLRP3 (green), and nuclei (DAPI; blue) (n=3 cochleae from three mice per group). Scale bars: 20 μm in A and B. (C) Quantification of SESN2- and NLRP3-positive cells among the three groups. (D) SESN2 expression in mice cochleae ( n = 3 cochleae from three mice per group). All experiments were replicated thrice. Data are presented as mean ± standard of the mean. * P < 0.05, ** P < 0.01, **** P < 0.0001 (one-way analysis of variance with Bonferroni’s multiple comparison test). Ctrl: Control; DAPI: 4,6-diamidino-2-phenylindole; NE: noise exposure; ns: not significant; TCP: tranylcypromine.

    Journal: Neural Regeneration Research

    Article Title: Tranylcypromine upregulates Sestrin 2 expression to ameliorate NLRP3-related noise-induced hearing loss

    doi: 10.4103/NRR.NRR-D-24-00130

    Figure Lengend Snippet: TCP increases SESN2 levels in NE mice cochleae. (A) Representative images of the medial cochlear turn in the control, NE+saline, and NE+TCP groups ( n = 3 cochleae from three mice per group). Hair cells were visualized by immunofluorescent staining for phalloidin (red), and the tissues were also stained for SESN2 (green), and nuclei (DAPI; blue). (B) Hair cells were visualized by immunofluorescent staining for phalloidin (red), and the tissues were also stained for NLRP3 (green), and nuclei (DAPI; blue) (n=3 cochleae from three mice per group). Scale bars: 20 μm in A and B. (C) Quantification of SESN2- and NLRP3-positive cells among the three groups. (D) SESN2 expression in mice cochleae ( n = 3 cochleae from three mice per group). All experiments were replicated thrice. Data are presented as mean ± standard of the mean. * P < 0.05, ** P < 0.01, **** P < 0.0001 (one-way analysis of variance with Bonferroni’s multiple comparison test). Ctrl: Control; DAPI: 4,6-diamidino-2-phenylindole; NE: noise exposure; ns: not significant; TCP: tranylcypromine.

    Article Snippet: First, the membranes were incubated overnight at 4°C with the following primary antibodies: mouse monoclonal anti-SESN2 (1:200, sc-393195, Santa Cruz Biotechnology, Santa Cruz, CA, USA), rabbit polyclonal anti-NLRP3 (1:400, NBP2-12446, Novus Biologicals, Centennial, CO, USA), rabbit polyclonal anti-LC3B (1:250, ab192890, Abcam, Cambridge, UK), rat monoclonal anti-LAMP1 (1:200, 14-1071-82, Invitrogen, Waltham, MA, USA), and mouse monoclonal 4-hydroxynonenal (4-HNE) (1:250, MA5-27570, Invitrogen).

    Techniques: Control, Saline, Staining, Expressing, Comparison

    NLRP3 expression in EOC and Kaplan-Meier survival analyses. ( a ) A box plot showed differential NLRP3 expression in normal and EOC samples. ( b ) Immunohistochemistry verified NLRP3 expression patterns in normal ovary (n=10) and EOC tissues (n=10) and their AODs; ( c ) Representative Western blot image showing NLRP3 protein expression in normal ovarian epithelial cells (IOSE80) and various EOC cell lines. Quantitative analysis of protein expression levels is shown below, with data represented as mean ± SD from three independent experiments. Symbols indicate statistically significant differences between IOSE80 and ovarian cancer cell lines. ( d – f ) Kaplan-Meier analysis revealed significant differences in patients’ DFS ( d ), PFS ( e ), and OS ( f ) between NLRP3 high and low expression groups in the training cohort. Data are shown as mean±SD; ns, no significance; ***, p < 0.001; ****, p < 0.0001.

    Journal: ImmunoTargets and Therapy

    Article Title: NLRP3 Inflammasome Upregulates PD-L1 in Ovarian Cancer and Contributes to an Immunosuppressive Microenvironment

    doi: 10.2147/ITT.S495564

    Figure Lengend Snippet: NLRP3 expression in EOC and Kaplan-Meier survival analyses. ( a ) A box plot showed differential NLRP3 expression in normal and EOC samples. ( b ) Immunohistochemistry verified NLRP3 expression patterns in normal ovary (n=10) and EOC tissues (n=10) and their AODs; ( c ) Representative Western blot image showing NLRP3 protein expression in normal ovarian epithelial cells (IOSE80) and various EOC cell lines. Quantitative analysis of protein expression levels is shown below, with data represented as mean ± SD from three independent experiments. Symbols indicate statistically significant differences between IOSE80 and ovarian cancer cell lines. ( d – f ) Kaplan-Meier analysis revealed significant differences in patients’ DFS ( d ), PFS ( e ), and OS ( f ) between NLRP3 high and low expression groups in the training cohort. Data are shown as mean±SD; ns, no significance; ***, p < 0.001; ****, p < 0.0001.

    Article Snippet: The following antibodies were utilized in this study: Rabbit Polyclonal anti-NLRP3 (CY5651), Rabbit Monoclonal anti- PD-L1 (CD274) Antibody (Abways, Shanghai, China; CY5980-20); Rabbit Polyclonal anti- Cleaved-Caspase-1(p20) (YC0022-20), Rabbit Polyclonal anti- IL-1β (YT5201), Rabbit Polyclonal anti- CXCL2(YT7842-20) (ImmunoWay Biotechnology, Plano, TX, USA).

    Techniques: Expressing, Immunohistochemistry, Western Blot

    Correlation between NLRP3 and immune infiltration and immune checkpoints. ( a ) The association of NLRP3 with stromal score, immune score, ESTIMATE score, and tumor purity was analyzed using the ESTIMATE algorithm. ( b ) The correlation between NLRP3 expression levels and signaling pathways enriched in EOC was analyzed by GSEA. ( c ) Scatter diagrams showed the correlation between NLRP3 and multiple immune checkpoints in EOC. ( d ) Representative immunofluorescence images from three independent experiments of NLRP3 and PD-L1 subcellular localization in the SKOV3 cell line. Nuclei were stained blue (DAPI), NLRP3 red, and PD-L1 green. ( e ) Pearson correlation analysis analyzed fluorescence co-localization of NLRP3 and PD-L1.

    Journal: ImmunoTargets and Therapy

    Article Title: NLRP3 Inflammasome Upregulates PD-L1 in Ovarian Cancer and Contributes to an Immunosuppressive Microenvironment

    doi: 10.2147/ITT.S495564

    Figure Lengend Snippet: Correlation between NLRP3 and immune infiltration and immune checkpoints. ( a ) The association of NLRP3 with stromal score, immune score, ESTIMATE score, and tumor purity was analyzed using the ESTIMATE algorithm. ( b ) The correlation between NLRP3 expression levels and signaling pathways enriched in EOC was analyzed by GSEA. ( c ) Scatter diagrams showed the correlation between NLRP3 and multiple immune checkpoints in EOC. ( d ) Representative immunofluorescence images from three independent experiments of NLRP3 and PD-L1 subcellular localization in the SKOV3 cell line. Nuclei were stained blue (DAPI), NLRP3 red, and PD-L1 green. ( e ) Pearson correlation analysis analyzed fluorescence co-localization of NLRP3 and PD-L1.

    Article Snippet: The following antibodies were utilized in this study: Rabbit Polyclonal anti-NLRP3 (CY5651), Rabbit Monoclonal anti- PD-L1 (CD274) Antibody (Abways, Shanghai, China; CY5980-20); Rabbit Polyclonal anti- Cleaved-Caspase-1(p20) (YC0022-20), Rabbit Polyclonal anti- IL-1β (YT5201), Rabbit Polyclonal anti- CXCL2(YT7842-20) (ImmunoWay Biotechnology, Plano, TX, USA).

    Techniques: Expressing, Immunofluorescence, Staining, Fluorescence

    NLRP3 inflammasome promotes PD-L1 expression and cell proliferation in SKOV3 cell line. ( a ) Inhibition of the NLRP3 inflammasome reduced the proliferation rate of SKOV3 cells, as analyzed by CCK-8 assays. ( b ) Western blot analysis was performed to measure IL-1β(p17), caspase-1 (p20), PD-L1, and NLRP3 under different treatments with LPS (6h) +ATP (1h) and/or MCC950 (1h) in the SKOV3 cell line. ( c and d ) Representative images of Colony formation(c) and TUNEL analysis(d) of SKOV3 cells under different treatment conditions were shown. Semi-quantitative analysis of colony numbers and apoptosis percentage were also included. Data were presented as mean ± SD from three independent experiments. ns, no significance, *, p < 0.05, ****, p < 0.0001. “#” represents each independent experiment.

    Journal: ImmunoTargets and Therapy

    Article Title: NLRP3 Inflammasome Upregulates PD-L1 in Ovarian Cancer and Contributes to an Immunosuppressive Microenvironment

    doi: 10.2147/ITT.S495564

    Figure Lengend Snippet: NLRP3 inflammasome promotes PD-L1 expression and cell proliferation in SKOV3 cell line. ( a ) Inhibition of the NLRP3 inflammasome reduced the proliferation rate of SKOV3 cells, as analyzed by CCK-8 assays. ( b ) Western blot analysis was performed to measure IL-1β(p17), caspase-1 (p20), PD-L1, and NLRP3 under different treatments with LPS (6h) +ATP (1h) and/or MCC950 (1h) in the SKOV3 cell line. ( c and d ) Representative images of Colony formation(c) and TUNEL analysis(d) of SKOV3 cells under different treatment conditions were shown. Semi-quantitative analysis of colony numbers and apoptosis percentage were also included. Data were presented as mean ± SD from three independent experiments. ns, no significance, *, p < 0.05, ****, p < 0.0001. “#” represents each independent experiment.

    Article Snippet: The following antibodies were utilized in this study: Rabbit Polyclonal anti-NLRP3 (CY5651), Rabbit Monoclonal anti- PD-L1 (CD274) Antibody (Abways, Shanghai, China; CY5980-20); Rabbit Polyclonal anti- Cleaved-Caspase-1(p20) (YC0022-20), Rabbit Polyclonal anti- IL-1β (YT5201), Rabbit Polyclonal anti- CXCL2(YT7842-20) (ImmunoWay Biotechnology, Plano, TX, USA).

    Techniques: Expressing, Inhibition, CCK-8 Assay, Western Blot, TUNEL Assay

    MCC950 application suppressed the activation of the NLRP3 inflammasome and slowed the progression of EOC in vivo. ( a ) Diagram illustrating the establishment of the murine model and the therapeutic intervention approaches. ( b – c ) Macroscopic tumor appearance across PBS-treated (n=3) and MCC950-treated (n=3) mice. ( d ) Tumor volume progression during the treatment period. ( e ) Comparison of the final tumor weight. (f-g) The levels of IL-1β (p17) in the serum ( f ) and tumor tissues ( g ) of control and MCC950-treated mice were quantified by ELISA ( f ) and Western blot ( g ), respectively. ( h – i ) The protein levels of caspase-1 (p20) (h) and PD-L1 (i) in the tumor tissues of the mice from control and MCC950-treated groups were assessed by Western blot. All numerical data are represented as the mean ± SD from at least three independent experimental repetitions. *, p < 0.05, **, p < 0.01, ****, p < 0.0001. n = biologically independent mouse samples. “#” represents each independent experiment.

    Journal: ImmunoTargets and Therapy

    Article Title: NLRP3 Inflammasome Upregulates PD-L1 in Ovarian Cancer and Contributes to an Immunosuppressive Microenvironment

    doi: 10.2147/ITT.S495564

    Figure Lengend Snippet: MCC950 application suppressed the activation of the NLRP3 inflammasome and slowed the progression of EOC in vivo. ( a ) Diagram illustrating the establishment of the murine model and the therapeutic intervention approaches. ( b – c ) Macroscopic tumor appearance across PBS-treated (n=3) and MCC950-treated (n=3) mice. ( d ) Tumor volume progression during the treatment period. ( e ) Comparison of the final tumor weight. (f-g) The levels of IL-1β (p17) in the serum ( f ) and tumor tissues ( g ) of control and MCC950-treated mice were quantified by ELISA ( f ) and Western blot ( g ), respectively. ( h – i ) The protein levels of caspase-1 (p20) (h) and PD-L1 (i) in the tumor tissues of the mice from control and MCC950-treated groups were assessed by Western blot. All numerical data are represented as the mean ± SD from at least three independent experimental repetitions. *, p < 0.05, **, p < 0.01, ****, p < 0.0001. n = biologically independent mouse samples. “#” represents each independent experiment.

    Article Snippet: The following antibodies were utilized in this study: Rabbit Polyclonal anti-NLRP3 (CY5651), Rabbit Monoclonal anti- PD-L1 (CD274) Antibody (Abways, Shanghai, China; CY5980-20); Rabbit Polyclonal anti- Cleaved-Caspase-1(p20) (YC0022-20), Rabbit Polyclonal anti- IL-1β (YT5201), Rabbit Polyclonal anti- CXCL2(YT7842-20) (ImmunoWay Biotechnology, Plano, TX, USA).

    Techniques: Activation Assay, In Vivo, Comparison, Control, Enzyme-linked Immunosorbent Assay, Western Blot

    Inhibition of the NLRP3 inflammasome leads to a decrease in immunosuppressive cells in murine ovarian cancer. ( a – c ) Flow cytometry analysis of PD-1 + CD4 + T cells ( a ), effector CD8 + ( b ) and CD4 + ( c ) T cells in spleens and tumors of mice treated with PBS (as control) or MCC950, and their quantification and statistical evaluation. ( d – f ) Representative flow cytometry graphs (left panel) and bar charts (right panel) demonstrating the frequencies and statistical evaluation of MDSCs ( d ), TAMs ( e ), and Tregs ( f ) in the tumor and spleen of control and MCC950-treated mice. ( g ) Analysis of CXCL2 protein expression in murine tumor tissues under different treatment conditions with and without MCC950 using Western blot. (All quantitative data are presented as mean ± SD from at least three biologically independent experiments, ns, no significance. **, p < 0.01, ***, p < 0.001, ****, p < 0.0001). “#” represents each independent experiment.

    Journal: ImmunoTargets and Therapy

    Article Title: NLRP3 Inflammasome Upregulates PD-L1 in Ovarian Cancer and Contributes to an Immunosuppressive Microenvironment

    doi: 10.2147/ITT.S495564

    Figure Lengend Snippet: Inhibition of the NLRP3 inflammasome leads to a decrease in immunosuppressive cells in murine ovarian cancer. ( a – c ) Flow cytometry analysis of PD-1 + CD4 + T cells ( a ), effector CD8 + ( b ) and CD4 + ( c ) T cells in spleens and tumors of mice treated with PBS (as control) or MCC950, and their quantification and statistical evaluation. ( d – f ) Representative flow cytometry graphs (left panel) and bar charts (right panel) demonstrating the frequencies and statistical evaluation of MDSCs ( d ), TAMs ( e ), and Tregs ( f ) in the tumor and spleen of control and MCC950-treated mice. ( g ) Analysis of CXCL2 protein expression in murine tumor tissues under different treatment conditions with and without MCC950 using Western blot. (All quantitative data are presented as mean ± SD from at least three biologically independent experiments, ns, no significance. **, p < 0.01, ***, p < 0.001, ****, p < 0.0001). “#” represents each independent experiment.

    Article Snippet: The following antibodies were utilized in this study: Rabbit Polyclonal anti-NLRP3 (CY5651), Rabbit Monoclonal anti- PD-L1 (CD274) Antibody (Abways, Shanghai, China; CY5980-20); Rabbit Polyclonal anti- Cleaved-Caspase-1(p20) (YC0022-20), Rabbit Polyclonal anti- IL-1β (YT5201), Rabbit Polyclonal anti- CXCL2(YT7842-20) (ImmunoWay Biotechnology, Plano, TX, USA).

    Techniques: Inhibition, Flow Cytometry, Control, Expressing, Western Blot

    In vitro characterization of a DNA vector expressing the multiepitopic CoV2-TMEP protein. ( A ) Schematic representation of the chimeric CoV2-TMEP multiepitopic vaccine targeting T cells. ( B ) Time-course expression of CoV2-TMEP protein. HEK-293T cells were transfected as detailed in the Materials and Methods section and CoV2-TMEP expression in pellets and supernatants (SN) was analyzed by Western blotting using the mouse monoclonal anti-FLAG antibody. ( C ) Subcellular localization of CoV2-TMEP protein. HeLa cells were transfected and processed for immunofluorescence microscopy as indicated in the Materials and Methods section. Green staining: mouse monoclonal anti-FLAG antibody followed by anti-mouse Alexa Fluor 488 antibody. Yellow arrows: CoV2-TMEP forming aggregates in magnified images (10×). ( D ) Analysis of NLRP3 induction and PARP cleavage. HEK-293T cells were transfected as described in the Materials and Methods section and the expression of NLRP3 (118 KDa; upper panel) and PARP (full-length: 116 KDa; cleaved: 89 KDa; lower panel) was analyzed by Western blotting using specific antibodies. ( E ) Subcellular colocalization of CoV2-TMEP protein with inflammasome. HeLa cells were transfected and processed for confocal microscopy as detailed in the Materials and Methods section. A mouse monoclonal anti-FLAG antibody and a rabbit polyclonal anti-NLRP3/NALP3 antibody followed by the corresponding secondary antibodies conjugated with Alexa Fluor 488 (green staining; FLAG detection) or Alexa Fluor 594 (red staining; NLRP3 detection) were used. Colocalization signal is indicated in white. Yellow arrows: Discrete colocalization signals in magnified images (5×). One representative optical section from three independent experiments is shown.

    Journal: Vaccines

    Article Title: B and T Cell Bi-Cistronic Multiepitopic Vaccine Induces Broad Immunogenicity and Provides Protection Against SARS-CoV-2

    doi: 10.3390/vaccines12111213

    Figure Lengend Snippet: In vitro characterization of a DNA vector expressing the multiepitopic CoV2-TMEP protein. ( A ) Schematic representation of the chimeric CoV2-TMEP multiepitopic vaccine targeting T cells. ( B ) Time-course expression of CoV2-TMEP protein. HEK-293T cells were transfected as detailed in the Materials and Methods section and CoV2-TMEP expression in pellets and supernatants (SN) was analyzed by Western blotting using the mouse monoclonal anti-FLAG antibody. ( C ) Subcellular localization of CoV2-TMEP protein. HeLa cells were transfected and processed for immunofluorescence microscopy as indicated in the Materials and Methods section. Green staining: mouse monoclonal anti-FLAG antibody followed by anti-mouse Alexa Fluor 488 antibody. Yellow arrows: CoV2-TMEP forming aggregates in magnified images (10×). ( D ) Analysis of NLRP3 induction and PARP cleavage. HEK-293T cells were transfected as described in the Materials and Methods section and the expression of NLRP3 (118 KDa; upper panel) and PARP (full-length: 116 KDa; cleaved: 89 KDa; lower panel) was analyzed by Western blotting using specific antibodies. ( E ) Subcellular colocalization of CoV2-TMEP protein with inflammasome. HeLa cells were transfected and processed for confocal microscopy as detailed in the Materials and Methods section. A mouse monoclonal anti-FLAG antibody and a rabbit polyclonal anti-NLRP3/NALP3 antibody followed by the corresponding secondary antibodies conjugated with Alexa Fluor 488 (green staining; FLAG detection) or Alexa Fluor 594 (red staining; NLRP3 detection) were used. Colocalization signal is indicated in white. Yellow arrows: Discrete colocalization signals in magnified images (5×). One representative optical section from three independent experiments is shown.

    Article Snippet: To evaluate the potential association between CoV2-TMEP and NLRP3, as well as the induction of PARP cleavage, monolayers of HEK-293T were transfected as described above, harvested at 24 hpt and analyzed by Western blotting using a rabbit polyclonal anti-NLRP3/NALP3 antibody (1:200; Cat. #NBP2-12446; Novus Biologicals, Centennial, CO, USA) or a rabbit polyclonal anti-PARP antibody (1:500; Cat. #9542; Cell Signaling, Danvers, MA, USA), followed by an HRP-conjugated anti-rabbit antibody (1:5000; Sigma-Aldrich).

    Techniques: In Vitro, Plasmid Preparation, Expressing, Transfection, Western Blot, Immunofluorescence, Microscopy, Staining, Confocal Microscopy