Review



tgn38  (Addgene inc)


Bioz Verified Symbol Addgene inc is a verified supplier  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
    • 93
      Buy from Supplier

    Structured Review

    Addgene inc tgn38
    a Conserved basic segments within the linker and FISNA region of NLRP3 homologs. b Immunoblot of non-localized and organelle-tethered NLRP3-YFP with respective NLRP3 mutants K127A/K128A/K129A/K130A (4xA). c – h Confocal images of NLRP3-KO iBMDMs stably transduced with organelle-tethered NLRP3-YFP (yellow) variants 12 h post priming with or without nigericin stimulation (Nig), stained for nuclei (Hoechst 33342, cyan) and plasma membrane (Cholera Toxin Subunit B, CTB, magenta). Scale bar, 10 µm. Following priming, 10 µM nigericin, 0.2 mg/ml SiO 2 or 20 ug/ml imiquimod (Imq) were added to cells, respectively, as indicated in the respective graph columns with legends above. Measurements of IL-1β were normalized to each respective non-mutated, organelle-enriched NLRP3-YFP: WT ( c ), Cb5 ( d ), <t>TGN38</t> ( e ), HRas ( f ), Rheb ( g ) or Omp25 ( h ) treated with canonical stimuli: nigericin, SiO 2 or imiquimod, respectively. EV, empty vector. Data represents the mean ± SEM of three independent experiments, except for TGN38, treated with SiO 2 in ( e ), which is the mean ± SEM of two independent experiments. The average value calculated from technical replicates within an individual experiment is presented as a single data point. P values were calculated using two-tailed unpaired t -test with Welch’s correction ( c – h ). Western blots ( b ) and microscopic images ( c – h ) are representative of three independent experiments. Panel ( a ) created in Biorender. Hafner Bratkovic, I. (2025) https://BioRender.com/99kdc99 .
    Tgn38, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/tgn38/product/Addgene inc
    Average 93 stars, based on 8 article reviews
    tgn38 - by Bioz Stars, 2026-03
    93/100 stars

    Images

    1) Product Images from "Clustering of NLRP3 induced by membrane or protein scaffolds promotes inflammasome assembly"

    Article Title: Clustering of NLRP3 induced by membrane or protein scaffolds promotes inflammasome assembly

    Journal: Nature Communications

    doi: 10.1038/s41467-025-60277-4

    a Conserved basic segments within the linker and FISNA region of NLRP3 homologs. b Immunoblot of non-localized and organelle-tethered NLRP3-YFP with respective NLRP3 mutants K127A/K128A/K129A/K130A (4xA). c – h Confocal images of NLRP3-KO iBMDMs stably transduced with organelle-tethered NLRP3-YFP (yellow) variants 12 h post priming with or without nigericin stimulation (Nig), stained for nuclei (Hoechst 33342, cyan) and plasma membrane (Cholera Toxin Subunit B, CTB, magenta). Scale bar, 10 µm. Following priming, 10 µM nigericin, 0.2 mg/ml SiO 2 or 20 ug/ml imiquimod (Imq) were added to cells, respectively, as indicated in the respective graph columns with legends above. Measurements of IL-1β were normalized to each respective non-mutated, organelle-enriched NLRP3-YFP: WT ( c ), Cb5 ( d ), TGN38 ( e ), HRas ( f ), Rheb ( g ) or Omp25 ( h ) treated with canonical stimuli: nigericin, SiO 2 or imiquimod, respectively. EV, empty vector. Data represents the mean ± SEM of three independent experiments, except for TGN38, treated with SiO 2 in ( e ), which is the mean ± SEM of two independent experiments. The average value calculated from technical replicates within an individual experiment is presented as a single data point. P values were calculated using two-tailed unpaired t -test with Welch’s correction ( c – h ). Western blots ( b ) and microscopic images ( c – h ) are representative of three independent experiments. Panel ( a ) created in Biorender. Hafner Bratkovic, I. (2025) https://BioRender.com/99kdc99 .
    Figure Legend Snippet: a Conserved basic segments within the linker and FISNA region of NLRP3 homologs. b Immunoblot of non-localized and organelle-tethered NLRP3-YFP with respective NLRP3 mutants K127A/K128A/K129A/K130A (4xA). c – h Confocal images of NLRP3-KO iBMDMs stably transduced with organelle-tethered NLRP3-YFP (yellow) variants 12 h post priming with or without nigericin stimulation (Nig), stained for nuclei (Hoechst 33342, cyan) and plasma membrane (Cholera Toxin Subunit B, CTB, magenta). Scale bar, 10 µm. Following priming, 10 µM nigericin, 0.2 mg/ml SiO 2 or 20 ug/ml imiquimod (Imq) were added to cells, respectively, as indicated in the respective graph columns with legends above. Measurements of IL-1β were normalized to each respective non-mutated, organelle-enriched NLRP3-YFP: WT ( c ), Cb5 ( d ), TGN38 ( e ), HRas ( f ), Rheb ( g ) or Omp25 ( h ) treated with canonical stimuli: nigericin, SiO 2 or imiquimod, respectively. EV, empty vector. Data represents the mean ± SEM of three independent experiments, except for TGN38, treated with SiO 2 in ( e ), which is the mean ± SEM of two independent experiments. The average value calculated from technical replicates within an individual experiment is presented as a single data point. P values were calculated using two-tailed unpaired t -test with Welch’s correction ( c – h ). Western blots ( b ) and microscopic images ( c – h ) are representative of three independent experiments. Panel ( a ) created in Biorender. Hafner Bratkovic, I. (2025) https://BioRender.com/99kdc99 .

    Techniques Used: Western Blot, Stable Transfection, Transduction, Staining, Clinical Proteomics, Membrane, Plasmid Preparation, Two Tailed Test



    Similar Products

    tgn38  (Bio-Rad)
    94
    Bio-Rad tgn38
    Tmem45b is essential for the calcium regulation of ER. (A) Representative immunostaining images show the colocalization between Tmem45b (green) and <t>TGN38</t> (red) in the DRG tissue sections. Scale bar, 20 μm. (B) The volcano plot illustrates the significant differences between WT and CKO mice (WT vs. cKO, n = 3 vs. 4), assessed based on the fold change (cKO/WT) and adjusted p-value (padj). Blue dots represent genes that are significantly downregulated (fold change <0.67; padj <0.1), red dots indicate significantly upregulated genes (fold change >1.5; padj <0.1). Genes represented by black dots show no significant differences between the groups. (C) qPCR analysis. (WT vs. cKO, n = 4 vs. 4). Columns represent the mean expression level of Atp2a1 mRNA normalized to Gapdh . (D) Western blot analysis. Experiments are repeated three times. (E) Schematic of calcium imaging in dissociated DRG neurons response to thapsigargin. (F) Representative calcium imaging response to thapsigargin (100 μM). (G) Statistical results. (WT vs. cKO, n = 90 vs. 72). (H) Schematic of calcium imaging in dissociated DRG neurons in response to caffeine (10 mM). (I) The heatmap shows the calcium activity of DRG neurons in response to two rounds of caffeine stimulations. Neuron numbers are shown on the left of the image. (J) Statistical results. (K) Schematic of calcium imaging in dissociated DRG neurons response to Trypsin (500 nM). (L, M) Representative calcium response to Trypsin. (N) Statistical results. (WT vs. cKO, n = 227 vs. 217). Two-tailed unpaired Student’s t test. DRG was obtained from at least three mice. Data are expressed as the Mean ± S.E.M. * P < 0.05, ** P < 0.01, *** P < 0.001.
    Tgn38, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/tgn38/product/Bio-Rad
    Average 94 stars, based on 1 article reviews
    tgn38 - by Bioz Stars, 2026-03
    94/100 stars
    		  Buy from Supplier
    mab6018 mouse anti beta actin proteintech 66009 1 ig mouse anti tgn38  (Proteintech)
    97
    Proteintech mab6018 mouse anti beta actin proteintech 66009 1 ig mouse anti tgn38
    Tmem45b is essential for the calcium regulation of ER. (A) Representative immunostaining images show the colocalization between Tmem45b (green) and <t>TGN38</t> (red) in the DRG tissue sections. Scale bar, 20 μm. (B) The volcano plot illustrates the significant differences between WT and CKO mice (WT vs. cKO, n = 3 vs. 4), assessed based on the fold change (cKO/WT) and adjusted p-value (padj). Blue dots represent genes that are significantly downregulated (fold change <0.67; padj <0.1), red dots indicate significantly upregulated genes (fold change >1.5; padj <0.1). Genes represented by black dots show no significant differences between the groups. (C) qPCR analysis. (WT vs. cKO, n = 4 vs. 4). Columns represent the mean expression level of Atp2a1 mRNA normalized to Gapdh . (D) Western blot analysis. Experiments are repeated three times. (E) Schematic of calcium imaging in dissociated DRG neurons response to thapsigargin. (F) Representative calcium imaging response to thapsigargin (100 μM). (G) Statistical results. (WT vs. cKO, n = 90 vs. 72). (H) Schematic of calcium imaging in dissociated DRG neurons in response to caffeine (10 mM). (I) The heatmap shows the calcium activity of DRG neurons in response to two rounds of caffeine stimulations. Neuron numbers are shown on the left of the image. (J) Statistical results. (K) Schematic of calcium imaging in dissociated DRG neurons response to Trypsin (500 nM). (L, M) Representative calcium response to Trypsin. (N) Statistical results. (WT vs. cKO, n = 227 vs. 217). Two-tailed unpaired Student’s t test. DRG was obtained from at least three mice. Data are expressed as the Mean ± S.E.M. * P < 0.05, ** P < 0.01, *** P < 0.001.
    Mab6018 Mouse Anti Beta Actin Proteintech 66009 1 Ig Mouse Anti Tgn38, supplied by Proteintech, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mab6018 mouse anti beta actin proteintech 66009 1 ig mouse anti tgn38/product/Proteintech
    Average 97 stars, based on 1 article reviews
    mab6018 mouse anti beta actin proteintech 66009 1 ig mouse anti tgn38 - by Bioz Stars, 2026-03
    97/100 stars
    		  Buy from Supplier
    sheep anti tgn38  (Bio-Rad)
    94
    Bio-Rad sheep anti tgn38
    Tmem45b is essential for the calcium regulation of ER. (A) Representative immunostaining images show the colocalization between Tmem45b (green) and <t>TGN38</t> (red) in the DRG tissue sections. Scale bar, 20 μm. (B) The volcano plot illustrates the significant differences between WT and CKO mice (WT vs. cKO, n = 3 vs. 4), assessed based on the fold change (cKO/WT) and adjusted p-value (padj). Blue dots represent genes that are significantly downregulated (fold change <0.67; padj <0.1), red dots indicate significantly upregulated genes (fold change >1.5; padj <0.1). Genes represented by black dots show no significant differences between the groups. (C) qPCR analysis. (WT vs. cKO, n = 4 vs. 4). Columns represent the mean expression level of Atp2a1 mRNA normalized to Gapdh . (D) Western blot analysis. Experiments are repeated three times. (E) Schematic of calcium imaging in dissociated DRG neurons response to thapsigargin. (F) Representative calcium imaging response to thapsigargin (100 μM). (G) Statistical results. (WT vs. cKO, n = 90 vs. 72). (H) Schematic of calcium imaging in dissociated DRG neurons in response to caffeine (10 mM). (I) The heatmap shows the calcium activity of DRG neurons in response to two rounds of caffeine stimulations. Neuron numbers are shown on the left of the image. (J) Statistical results. (K) Schematic of calcium imaging in dissociated DRG neurons response to Trypsin (500 nM). (L, M) Representative calcium response to Trypsin. (N) Statistical results. (WT vs. cKO, n = 227 vs. 217). Two-tailed unpaired Student’s t test. DRG was obtained from at least three mice. Data are expressed as the Mean ± S.E.M. * P < 0.05, ** P < 0.01, *** P < 0.001.
    Sheep Anti Tgn38, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sheep anti tgn38/product/Bio-Rad
    Average 94 stars, based on 1 article reviews
    sheep anti tgn38 - by Bioz Stars, 2026-03
    94/100 stars
    		  Buy from Supplier
    antibody against mouse tgn38 ahp499g  (Bio-Rad)
    94
    Bio-Rad antibody against mouse tgn38 ahp499g
    Tmem45b is essential for the calcium regulation of ER. (A) Representative immunostaining images show the colocalization between Tmem45b (green) and <t>TGN38</t> (red) in the DRG tissue sections. Scale bar, 20 μm. (B) The volcano plot illustrates the significant differences between WT and CKO mice (WT vs. cKO, n = 3 vs. 4), assessed based on the fold change (cKO/WT) and adjusted p-value (padj). Blue dots represent genes that are significantly downregulated (fold change <0.67; padj <0.1), red dots indicate significantly upregulated genes (fold change >1.5; padj <0.1). Genes represented by black dots show no significant differences between the groups. (C) qPCR analysis. (WT vs. cKO, n = 4 vs. 4). Columns represent the mean expression level of Atp2a1 mRNA normalized to Gapdh . (D) Western blot analysis. Experiments are repeated three times. (E) Schematic of calcium imaging in dissociated DRG neurons response to thapsigargin. (F) Representative calcium imaging response to thapsigargin (100 μM). (G) Statistical results. (WT vs. cKO, n = 90 vs. 72). (H) Schematic of calcium imaging in dissociated DRG neurons in response to caffeine (10 mM). (I) The heatmap shows the calcium activity of DRG neurons in response to two rounds of caffeine stimulations. Neuron numbers are shown on the left of the image. (J) Statistical results. (K) Schematic of calcium imaging in dissociated DRG neurons response to Trypsin (500 nM). (L, M) Representative calcium response to Trypsin. (N) Statistical results. (WT vs. cKO, n = 227 vs. 217). Two-tailed unpaired Student’s t test. DRG was obtained from at least three mice. Data are expressed as the Mean ± S.E.M. * P < 0.05, ** P < 0.01, *** P < 0.001.
    Antibody Against Mouse Tgn38 Ahp499g, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/antibody against mouse tgn38 ahp499g/product/Bio-Rad
    Average 94 stars, based on 1 article reviews
    antibody against mouse tgn38 ahp499g - by Bioz Stars, 2026-03
    94/100 stars
    		  Buy from Supplier
    anti-tgn38 nbp1-03495  (Novus Biologicals)
    90
    Novus Biologicals anti-tgn38 nbp1-03495
    Tmem45b is essential for the calcium regulation of ER. (A) Representative immunostaining images show the colocalization between Tmem45b (green) and <t>TGN38</t> (red) in the DRG tissue sections. Scale bar, 20 μm. (B) The volcano plot illustrates the significant differences between WT and CKO mice (WT vs. cKO, n = 3 vs. 4), assessed based on the fold change (cKO/WT) and adjusted p-value (padj). Blue dots represent genes that are significantly downregulated (fold change <0.67; padj <0.1), red dots indicate significantly upregulated genes (fold change >1.5; padj <0.1). Genes represented by black dots show no significant differences between the groups. (C) qPCR analysis. (WT vs. cKO, n = 4 vs. 4). Columns represent the mean expression level of Atp2a1 mRNA normalized to Gapdh . (D) Western blot analysis. Experiments are repeated three times. (E) Schematic of calcium imaging in dissociated DRG neurons response to thapsigargin. (F) Representative calcium imaging response to thapsigargin (100 μM). (G) Statistical results. (WT vs. cKO, n = 90 vs. 72). (H) Schematic of calcium imaging in dissociated DRG neurons in response to caffeine (10 mM). (I) The heatmap shows the calcium activity of DRG neurons in response to two rounds of caffeine stimulations. Neuron numbers are shown on the left of the image. (J) Statistical results. (K) Schematic of calcium imaging in dissociated DRG neurons response to Trypsin (500 nM). (L, M) Representative calcium response to Trypsin. (N) Statistical results. (WT vs. cKO, n = 227 vs. 217). Two-tailed unpaired Student’s t test. DRG was obtained from at least three mice. Data are expressed as the Mean ± S.E.M. * P < 0.05, ** P < 0.01, *** P < 0.001.
    Anti Tgn38 Nbp1 03495, supplied by Novus Biologicals, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti-tgn38 nbp1-03495/product/Novus Biologicals
    Average 90 stars, based on 1 article reviews
    anti-tgn38 nbp1-03495 - by Bioz Stars, 2026-03
    90/100 stars
    		  Buy from Supplier
    tgn38  (Addgene inc)
    93
    Addgene inc tgn38
    a Conserved basic segments within the linker and FISNA region of NLRP3 homologs. b Immunoblot of non-localized and organelle-tethered NLRP3-YFP with respective NLRP3 mutants K127A/K128A/K129A/K130A (4xA). c – h Confocal images of NLRP3-KO iBMDMs stably transduced with organelle-tethered NLRP3-YFP (yellow) variants 12 h post priming with or without nigericin stimulation (Nig), stained for nuclei (Hoechst 33342, cyan) and plasma membrane (Cholera Toxin Subunit B, CTB, magenta). Scale bar, 10 µm. Following priming, 10 µM nigericin, 0.2 mg/ml SiO 2 or 20 ug/ml imiquimod (Imq) were added to cells, respectively, as indicated in the respective graph columns with legends above. Measurements of IL-1β were normalized to each respective non-mutated, organelle-enriched NLRP3-YFP: WT ( c ), Cb5 ( d ), <t>TGN38</t> ( e ), HRas ( f ), Rheb ( g ) or Omp25 ( h ) treated with canonical stimuli: nigericin, SiO 2 or imiquimod, respectively. EV, empty vector. Data represents the mean ± SEM of three independent experiments, except for TGN38, treated with SiO 2 in ( e ), which is the mean ± SEM of two independent experiments. The average value calculated from technical replicates within an individual experiment is presented as a single data point. P values were calculated using two-tailed unpaired t -test with Welch’s correction ( c – h ). Western blots ( b ) and microscopic images ( c – h ) are representative of three independent experiments. Panel ( a ) created in Biorender. Hafner Bratkovic, I. (2025) https://BioRender.com/99kdc99 .
    Tgn38, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/tgn38/product/Addgene inc
    Average 93 stars, based on 1 article reviews
    tgn38 - by Bioz Stars, 2026-03
    93/100 stars
    		  Buy from Supplier
    mouse anti tgn38 46  (Santa Cruz Biotechnology)
    94
    Santa Cruz Biotechnology mouse anti tgn38 46
    a Conserved basic segments within the linker and FISNA region of NLRP3 homologs. b Immunoblot of non-localized and organelle-tethered NLRP3-YFP with respective NLRP3 mutants K127A/K128A/K129A/K130A (4xA). c – h Confocal images of NLRP3-KO iBMDMs stably transduced with organelle-tethered NLRP3-YFP (yellow) variants 12 h post priming with or without nigericin stimulation (Nig), stained for nuclei (Hoechst 33342, cyan) and plasma membrane (Cholera Toxin Subunit B, CTB, magenta). Scale bar, 10 µm. Following priming, 10 µM nigericin, 0.2 mg/ml SiO 2 or 20 ug/ml imiquimod (Imq) were added to cells, respectively, as indicated in the respective graph columns with legends above. Measurements of IL-1β were normalized to each respective non-mutated, organelle-enriched NLRP3-YFP: WT ( c ), Cb5 ( d ), <t>TGN38</t> ( e ), HRas ( f ), Rheb ( g ) or Omp25 ( h ) treated with canonical stimuli: nigericin, SiO 2 or imiquimod, respectively. EV, empty vector. Data represents the mean ± SEM of three independent experiments, except for TGN38, treated with SiO 2 in ( e ), which is the mean ± SEM of two independent experiments. The average value calculated from technical replicates within an individual experiment is presented as a single data point. P values were calculated using two-tailed unpaired t -test with Welch’s correction ( c – h ). Western blots ( b ) and microscopic images ( c – h ) are representative of three independent experiments. Panel ( a ) created in Biorender. Hafner Bratkovic, I. (2025) https://BioRender.com/99kdc99 .
    Mouse Anti Tgn38 46, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mouse anti tgn38 46/product/Santa Cruz Biotechnology
    Average 94 stars, based on 1 article reviews
    mouse anti tgn38 46 - by Bioz Stars, 2026-03
    94/100 stars
    		  Buy from Supplier
    34 tgn38 santa cruz sc 166594  (Santa Cruz Biotechnology)
    94
    Santa Cruz Biotechnology 34 tgn38 santa cruz sc 166594
    a Conserved basic segments within the linker and FISNA region of NLRP3 homologs. b Immunoblot of non-localized and organelle-tethered NLRP3-YFP with respective NLRP3 mutants K127A/K128A/K129A/K130A (4xA). c – h Confocal images of NLRP3-KO iBMDMs stably transduced with organelle-tethered NLRP3-YFP (yellow) variants 12 h post priming with or without nigericin stimulation (Nig), stained for nuclei (Hoechst 33342, cyan) and plasma membrane (Cholera Toxin Subunit B, CTB, magenta). Scale bar, 10 µm. Following priming, 10 µM nigericin, 0.2 mg/ml SiO 2 or 20 ug/ml imiquimod (Imq) were added to cells, respectively, as indicated in the respective graph columns with legends above. Measurements of IL-1β were normalized to each respective non-mutated, organelle-enriched NLRP3-YFP: WT ( c ), Cb5 ( d ), <t>TGN38</t> ( e ), HRas ( f ), Rheb ( g ) or Omp25 ( h ) treated with canonical stimuli: nigericin, SiO 2 or imiquimod, respectively. EV, empty vector. Data represents the mean ± SEM of three independent experiments, except for TGN38, treated with SiO 2 in ( e ), which is the mean ± SEM of two independent experiments. The average value calculated from technical replicates within an individual experiment is presented as a single data point. P values were calculated using two-tailed unpaired t -test with Welch’s correction ( c – h ). Western blots ( b ) and microscopic images ( c – h ) are representative of three independent experiments. Panel ( a ) created in Biorender. Hafner Bratkovic, I. (2025) https://BioRender.com/99kdc99 .
    34 Tgn38 Santa Cruz Sc 166594, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/34 tgn38 santa cruz sc 166594/product/Santa Cruz Biotechnology
    Average 94 stars, based on 1 article reviews
    34 tgn38 santa cruz sc 166594 - by Bioz Stars, 2026-03
    94/100 stars
    		  Buy from Supplier

    Image Search Results


    Tmem45b is essential for the calcium regulation of ER. (A) Representative immunostaining images show the colocalization between Tmem45b (green) and TGN38 (red) in the DRG tissue sections. Scale bar, 20 μm. (B) The volcano plot illustrates the significant differences between WT and CKO mice (WT vs. cKO, n = 3 vs. 4), assessed based on the fold change (cKO/WT) and adjusted p-value (padj). Blue dots represent genes that are significantly downregulated (fold change <0.67; padj <0.1), red dots indicate significantly upregulated genes (fold change >1.5; padj <0.1). Genes represented by black dots show no significant differences between the groups. (C) qPCR analysis. (WT vs. cKO, n = 4 vs. 4). Columns represent the mean expression level of Atp2a1 mRNA normalized to Gapdh . (D) Western blot analysis. Experiments are repeated three times. (E) Schematic of calcium imaging in dissociated DRG neurons response to thapsigargin. (F) Representative calcium imaging response to thapsigargin (100 μM). (G) Statistical results. (WT vs. cKO, n = 90 vs. 72). (H) Schematic of calcium imaging in dissociated DRG neurons in response to caffeine (10 mM). (I) The heatmap shows the calcium activity of DRG neurons in response to two rounds of caffeine stimulations. Neuron numbers are shown on the left of the image. (J) Statistical results. (K) Schematic of calcium imaging in dissociated DRG neurons response to Trypsin (500 nM). (L, M) Representative calcium response to Trypsin. (N) Statistical results. (WT vs. cKO, n = 227 vs. 217). Two-tailed unpaired Student’s t test. DRG was obtained from at least three mice. Data are expressed as the Mean ± S.E.M. * P < 0.05, ** P < 0.01, *** P < 0.001.

    Journal: Frontiers in Physiology

    Article Title: Tmem45b modulates itch via endoplasmic reticulum calcium regulation

    doi: 10.3389/fphys.2025.1708686

    Figure Lengend Snippet: Tmem45b is essential for the calcium regulation of ER. (A) Representative immunostaining images show the colocalization between Tmem45b (green) and TGN38 (red) in the DRG tissue sections. Scale bar, 20 μm. (B) The volcano plot illustrates the significant differences between WT and CKO mice (WT vs. cKO, n = 3 vs. 4), assessed based on the fold change (cKO/WT) and adjusted p-value (padj). Blue dots represent genes that are significantly downregulated (fold change <0.67; padj <0.1), red dots indicate significantly upregulated genes (fold change >1.5; padj <0.1). Genes represented by black dots show no significant differences between the groups. (C) qPCR analysis. (WT vs. cKO, n = 4 vs. 4). Columns represent the mean expression level of Atp2a1 mRNA normalized to Gapdh . (D) Western blot analysis. Experiments are repeated three times. (E) Schematic of calcium imaging in dissociated DRG neurons response to thapsigargin. (F) Representative calcium imaging response to thapsigargin (100 μM). (G) Statistical results. (WT vs. cKO, n = 90 vs. 72). (H) Schematic of calcium imaging in dissociated DRG neurons in response to caffeine (10 mM). (I) The heatmap shows the calcium activity of DRG neurons in response to two rounds of caffeine stimulations. Neuron numbers are shown on the left of the image. (J) Statistical results. (K) Schematic of calcium imaging in dissociated DRG neurons response to Trypsin (500 nM). (L, M) Representative calcium response to Trypsin. (N) Statistical results. (WT vs. cKO, n = 227 vs. 217). Two-tailed unpaired Student’s t test. DRG was obtained from at least three mice. Data are expressed as the Mean ± S.E.M. * P < 0.05, ** P < 0.01, *** P < 0.001.

    Article Snippet: Antibodies used in this study: GM130 (BD, 610822), Th (Millipore, AB1542), IB4 (Vector, FL-1201-.5), Tuj1 (Starter, SDT-251-28), CGRP (Dia Sorin, 24112), NF200 (CST, 2836S), PDI (Santa Cruz, SC-20132), Calnexin (Abcam, ab112995), Serca1 (Proteintech, 22361-1-AP), mitochondria-tracer (Beyotime, C1048), TGN38 (Bio-Rad, AHP499G), GFAP (Millipore, MAB3402), IBA1 (Abcam, ab5076).

    Techniques: Immunostaining, Expressing, Western Blot, Imaging, Activity Assay, Two Tailed Test

    a Conserved basic segments within the linker and FISNA region of NLRP3 homologs. b Immunoblot of non-localized and organelle-tethered NLRP3-YFP with respective NLRP3 mutants K127A/K128A/K129A/K130A (4xA). c – h Confocal images of NLRP3-KO iBMDMs stably transduced with organelle-tethered NLRP3-YFP (yellow) variants 12 h post priming with or without nigericin stimulation (Nig), stained for nuclei (Hoechst 33342, cyan) and plasma membrane (Cholera Toxin Subunit B, CTB, magenta). Scale bar, 10 µm. Following priming, 10 µM nigericin, 0.2 mg/ml SiO 2 or 20 ug/ml imiquimod (Imq) were added to cells, respectively, as indicated in the respective graph columns with legends above. Measurements of IL-1β were normalized to each respective non-mutated, organelle-enriched NLRP3-YFP: WT ( c ), Cb5 ( d ), TGN38 ( e ), HRas ( f ), Rheb ( g ) or Omp25 ( h ) treated with canonical stimuli: nigericin, SiO 2 or imiquimod, respectively. EV, empty vector. Data represents the mean ± SEM of three independent experiments, except for TGN38, treated with SiO 2 in ( e ), which is the mean ± SEM of two independent experiments. The average value calculated from technical replicates within an individual experiment is presented as a single data point. P values were calculated using two-tailed unpaired t -test with Welch’s correction ( c – h ). Western blots ( b ) and microscopic images ( c – h ) are representative of three independent experiments. Panel ( a ) created in Biorender. Hafner Bratkovic, I. (2025) https://BioRender.com/99kdc99 .

    Journal: Nature Communications

    Article Title: Clustering of NLRP3 induced by membrane or protein scaffolds promotes inflammasome assembly

    doi: 10.1038/s41467-025-60277-4

    Figure Lengend Snippet: a Conserved basic segments within the linker and FISNA region of NLRP3 homologs. b Immunoblot of non-localized and organelle-tethered NLRP3-YFP with respective NLRP3 mutants K127A/K128A/K129A/K130A (4xA). c – h Confocal images of NLRP3-KO iBMDMs stably transduced with organelle-tethered NLRP3-YFP (yellow) variants 12 h post priming with or without nigericin stimulation (Nig), stained for nuclei (Hoechst 33342, cyan) and plasma membrane (Cholera Toxin Subunit B, CTB, magenta). Scale bar, 10 µm. Following priming, 10 µM nigericin, 0.2 mg/ml SiO 2 or 20 ug/ml imiquimod (Imq) were added to cells, respectively, as indicated in the respective graph columns with legends above. Measurements of IL-1β were normalized to each respective non-mutated, organelle-enriched NLRP3-YFP: WT ( c ), Cb5 ( d ), TGN38 ( e ), HRas ( f ), Rheb ( g ) or Omp25 ( h ) treated with canonical stimuli: nigericin, SiO 2 or imiquimod, respectively. EV, empty vector. Data represents the mean ± SEM of three independent experiments, except for TGN38, treated with SiO 2 in ( e ), which is the mean ± SEM of two independent experiments. The average value calculated from technical replicates within an individual experiment is presented as a single data point. P values were calculated using two-tailed unpaired t -test with Welch’s correction ( c – h ). Western blots ( b ) and microscopic images ( c – h ) are representative of three independent experiments. Panel ( a ) created in Biorender. Hafner Bratkovic, I. (2025) https://BioRender.com/99kdc99 .

    Article Snippet: Localization sequences were amplified from plasmids encoding Hras (a gift from D. Sabatini, Addgene 26637 ), Tmem192 (a gift from D. Sabatini, Addgene 102930 ), Rheb (a gift from D. Sabatini, Addgene 26634 ), TGN38 (a gift from J. Lippincott-Schwartz, Addgene 128148), GOLGB1 (a gift from D. Gadella, Addgene 67903 ), Omp25 (a gift from D. Sabatini, Addgene 26638 ), PEX3 (a gift from J. Boehm & W. Hahn & D. Root, Addgene 81779 ), PEX26 (a gift from D. Sabatini, Addgene 139054 ) and PACT (a gift from C. Norden, Addgene 105954 ).

    Techniques: Western Blot, Stable Transfection, Transduction, Staining, Clinical Proteomics, Membrane, Plasmid Preparation, Two Tailed Test