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anti synaptopodin catalog no sc 515842 antibody  (Santa Cruz Biotechnology)


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    Santa Cruz Biotechnology anti synaptopodin catalog no sc 515842 antibody
    Anti Synaptopodin Catalog No Sc 515842 Antibody, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 95/100, based on 372 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/synaptopodin/Synaptopodin+Antibody/pm41866523-358-24-32
    Average 95 stars, based on 372 article reviews
    anti synaptopodin catalog no sc 515842 antibody - by Bioz Stars, 2026-10
    95/100 stars

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    Related Articles

    Blocking Assay:

    Article Title: INPP5D/SHIP1-mediated immunometabolic remodeling of renal monocytes in idiopathic membranous nephropathy
    Article Snippet: .. After blocking with normal serum, sections were incubated with primary antibodies against CD163 (Abmart, TD8235S, China) and synaptopodin (Santa Cruz, sc-515842, US) followed by fluorescently labeled secondary antibodies. ..

    Incubation:

    Article Title: INPP5D/SHIP1-mediated immunometabolic remodeling of renal monocytes in idiopathic membranous nephropathy
    Article Snippet: .. After blocking with normal serum, sections were incubated with primary antibodies against CD163 (Abmart, TD8235S, China) and synaptopodin (Santa Cruz, sc-515842, US) followed by fluorescently labeled secondary antibodies. ..

    Article Title: Extracellular Vesicles Facilitate the Crosstalk Between High Glucose-Stimulated Mesangial Cells and Healthy Podocytes to Mediate Injury Responses
    Article Snippet: Podocytes were grown on Lab-Tek® chamber slides (Thermo Fisher Scientific) and treated with HG (HG), HG-EVs, or C-EVs for 24 h. Cells were fixed with 3.5% formaldehyde for 15 min at room temperature, permeabilized with 0.2% Triton X-100 for 10 min, and blocked in PBS containing 0.5% bovine serum albumin (BSA) for 1h. .. Cells were incubated overnight at 4 ◦C with primary antibodies against nephrin (Abcam, 1:100), podocin (Abcam, 1:100), synaptopodin (Santa Cruz Biotechnology, 1:100), or vimentin (Thermo Fisher Scientific 1:100), followed by Alexa Fluor–conjugated secondary antibodies (488 or 594; Thermo Fisher Scientific, 1:1000) for 60 min at room temperature in the dark. .. Actin cytoskeleton organization was assessed using Alexa Fluor Plus 647-conjugated phalloidin (Invitrogen), following the manufacturer’s instructions.

    Article Title: Hypoxia-inducible factor 2α overexpression in podocytes ameliorates lipid metabolism disorders in diabetic kidney disease by inhibiting S1P
    Article Snippet: .. Nephrin (1:100, Santa Cruz, California, USA) and synaptopodin (1:100, Santa Cruz, California, USA) primary antibodies were incubated with podocytes overnight at 4 °C, followed by incubation with FITC-coupled secondary antibodies (1:100, Zsbio, Beijing, China) at room temperature for 1 h. Subsequently, podocytes were incubated with DAPI (Solarbio, Beijing, China) for 5 min. ..

    Article Title: Dual blockade of DPP-4 and CXCL12/CXCR4 axes synergistically protects podocytes in lupus nephritis
    Article Snippet: Tissue sections were fixed, permeabilized, and blocked with 3% BSA ( GC305010 , Servicebio). .. Sections were incubated overnight at 4 °C with primary antibodies against SDF1 (ab25117, Abcam), synaptopodin (sc-515842, SANTA CRUZ BIOTECHNOLOGY), IgG (ab172730, Abcam), NOX4 (ABC459, Sigma-Aldrich), and WT1 (ab89901, Abcam). ..

    Article Title: Extracellular Vesicles Facilitate the Crosstalk Between High Glucose-Stimulated Mesangial Cells and Healthy Podocytes to Mediate Injury Responses
    Article Snippet: Podocytes were grown on Lab-Tek ® chamber slides (Thermo Fisher Scientific) and treated with HG (HG), HG-EVs, or C-EVs for 24 h. Cells were fixed with 3.5% formaldehyde for 15 min at room temperature, permeabilized with 0.2% Triton X-100 for 10 min, and blocked in PBS containing 0.5% bovine serum albumin (BSA) for 1h. .. Cells were incubated overnight at 4 °C with primary antibodies against nephrin (Abcam, 1:100), podocin (Abcam, 1:100), synaptopodin (Santa Cruz Biotechnology, 1:100), or vimentin (Thermo Fisher Scientific 1:100), followed by Alexa Fluor–conjugated secondary antibodies (488 or 594; Thermo Fisher Scientific, 1:1000) for 60 min at room temperature in the dark. .. Actin cytoskeleton organization was assessed using Alexa Fluor Plus 647-conjugated phalloidin (Invitrogen), following the manufacturer’s instructions.

    Labeling:

    Article Title: INPP5D/SHIP1-mediated immunometabolic remodeling of renal monocytes in idiopathic membranous nephropathy
    Article Snippet: .. After blocking with normal serum, sections were incubated with primary antibodies against CD163 (Abmart, TD8235S, China) and synaptopodin (Santa Cruz, sc-515842, US) followed by fluorescently labeled secondary antibodies. ..

    Expressing:

    Article Title: Identification and Validation of miR-206-3p Targeting WT-1 Promotes Membranous Nephropathy Through a Comprehensive Bioinformatics and Machine Learning Algorithm.
    Article Snippet: Membranous nephropathy (MN), a major cause of endstage renal disease, has limited therapeutics due to unclear targets.. MicroRNAs (miRs) play critical roles in various diseases, but their role in MN remains unknown.. This study utilizes a series of public databases, bioinformatics tools, and validation methods to identify key miRs and molecular mechanisms involved in MN.

    Western Blot:

    Article Title: From Wild Vegetable to Renal Protector: The Therapeutic Potential of Aralia elata (Miq.) Seem
    Article Snippet: .. Besides, antibodies used for western blot analysis, immunohistochemical staining, and immunofluorescence staining included GPX4 (ab125066, Abcam, UK), ferritin heavy chain (FTH) (ab183781, Abcam, UK), ferroportin1 (FPN1) (26601–1‐AP, Proteintech, USA), SLC7A11 (ab307601, Abcam, UK), NPHS2 (ab181143, Abcam, UK), Synaptopodin (sc‐515842, Santa Cruz, USA), β‐actin (4970S, Cell Signaling Technology, USA), Anti‐rabbit immunoglobulin G (7074S, Cell Signaling Technology, USA). .. Twelve male BALB/c mice weighing 25 ± 5 g were bought from JOINN Laboratory, Suzhou, China (NO: 202236274) and adaptively fed for 1 week before the experiment.

    Immunohistochemical staining:

    Article Title: From Wild Vegetable to Renal Protector: The Therapeutic Potential of Aralia elata (Miq.) Seem
    Article Snippet: .. Besides, antibodies used for western blot analysis, immunohistochemical staining, and immunofluorescence staining included GPX4 (ab125066, Abcam, UK), ferritin heavy chain (FTH) (ab183781, Abcam, UK), ferroportin1 (FPN1) (26601–1‐AP, Proteintech, USA), SLC7A11 (ab307601, Abcam, UK), NPHS2 (ab181143, Abcam, UK), Synaptopodin (sc‐515842, Santa Cruz, USA), β‐actin (4970S, Cell Signaling Technology, USA), Anti‐rabbit immunoglobulin G (7074S, Cell Signaling Technology, USA). .. Twelve male BALB/c mice weighing 25 ± 5 g were bought from JOINN Laboratory, Suzhou, China (NO: 202236274) and adaptively fed for 1 week before the experiment.

    Staining:

    Article Title: From Wild Vegetable to Renal Protector: The Therapeutic Potential of Aralia elata (Miq.) Seem
    Article Snippet: .. Besides, antibodies used for western blot analysis, immunohistochemical staining, and immunofluorescence staining included GPX4 (ab125066, Abcam, UK), ferritin heavy chain (FTH) (ab183781, Abcam, UK), ferroportin1 (FPN1) (26601–1‐AP, Proteintech, USA), SLC7A11 (ab307601, Abcam, UK), NPHS2 (ab181143, Abcam, UK), Synaptopodin (sc‐515842, Santa Cruz, USA), β‐actin (4970S, Cell Signaling Technology, USA), Anti‐rabbit immunoglobulin G (7074S, Cell Signaling Technology, USA). .. Twelve male BALB/c mice weighing 25 ± 5 g were bought from JOINN Laboratory, Suzhou, China (NO: 202236274) and adaptively fed for 1 week before the experiment.

    Immunofluorescence:

    Article Title: From Wild Vegetable to Renal Protector: The Therapeutic Potential of Aralia elata (Miq.) Seem
    Article Snippet: .. Besides, antibodies used for western blot analysis, immunohistochemical staining, and immunofluorescence staining included GPX4 (ab125066, Abcam, UK), ferritin heavy chain (FTH) (ab183781, Abcam, UK), ferroportin1 (FPN1) (26601–1‐AP, Proteintech, USA), SLC7A11 (ab307601, Abcam, UK), NPHS2 (ab181143, Abcam, UK), Synaptopodin (sc‐515842, Santa Cruz, USA), β‐actin (4970S, Cell Signaling Technology, USA), Anti‐rabbit immunoglobulin G (7074S, Cell Signaling Technology, USA). .. Twelve male BALB/c mice weighing 25 ± 5 g were bought from JOINN Laboratory, Suzhou, China (NO: 202236274) and adaptively fed for 1 week before the experiment.

    other:

    Article Title: Unveiling the TrkA-p35/CDK5 axis: a novel therapeutic target in diabetic kidney disease
    Article Snippet: Antibodies against ERK (Cat. No. sc-514302), phospho-ERK (Cat. No. sc-7383), and Synaptopodin (Cat. No. sc-515842) were purchased from Santa Cruz Biotechnology (Dallas, TX, USA).



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    Podocyte-specific TLR4 deletion mitigated STZ-induced DKD, podocyte injury, and proteinuria in mice. (A) Schematic description of the strategy for generating STZ-induced diabetic mouse model in NPHS2 -Cre; TLR4 fl/fl ( TLR4 CKO) mice and in TLR4 fl/fl mice (CTL). (B) Monthly monitoring of plasma glucose levels in mice. Data are presented as mean ± SD. ** P < 0.01 vs. CTL-NC; ## P < 0.01 vs. CKO-NC. (C) Monthly monitoring of body weight in mice. Data are presented as mean ± SD. ** P < 0.01 vs. CTL-NC; # P < 0.05 vs. CKO-NC; # # P < 0.01 vs. CKO-NC; # ### P < 0.01 vs. CTL-STZ. (D) Representative images of PAS and MASSON staining of the glomeruli in mice at 12 and 16 weeks. Scale bar, 20 μm ( n = 8). (E) Representative images of HE staining of the kidney in mice at 12 and 16 weeks. Scale bar, 50 μm ( n = 8). (F) Representative IF images of the expression of SD proteins p-cadherin and <t>synaptopodin</t> in glomeruli at 12 and 16 weeks. Scale bar, 20 μm ( n = 8). (G) The expression of a mesenchymal marker desmin was detected in renal tissues from mice at 12 and 16 weeks by western blot analysis. P values were determined by one-way ANOVA and data are presented as mean ± SD. **** P < 0.01; # P < 0.05; ## P < 0.05 ( n = 8). (H) Representative images of TEM in podocyte ultrastructural alterations in mice. FPs effacement (blue arrow), ER dilation (red arrow) and GBM thickness were observed in podocytes in STZ-induced CTL diabetic mice, and were attenuated in TLR4 CKO diabetic mice at 16 weeks. Scale bar, 1 μm ( n = 8). (I) Determination of proteinuria in mice. Data are presented as mean ± SD. ** P < 0.01 vs. CTL-NC; # P < 0.05 vs. CKO-NC; # # P < 0.01 vs. CKO-NC; # ## P < 0.05 vs. CTL-STZ; # ### P < 0.01 vs. CTL-STZ.
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    anti synaptopodin catalog no sc 515842 antibody  (Santa Cruz Biotechnology)
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    Podocyte-specific TLR4 deletion mitigated STZ-induced DKD, podocyte injury, and proteinuria in mice. (A) Schematic description of the strategy for generating STZ-induced diabetic mouse model in NPHS2 -Cre; TLR4 fl/fl ( TLR4 CKO) mice and in TLR4 fl/fl mice (CTL). (B) Monthly monitoring of plasma glucose levels in mice. Data are presented as mean ± SD. ** P < 0.01 vs. CTL-NC; ## P < 0.01 vs. CKO-NC. (C) Monthly monitoring of body weight in mice. Data are presented as mean ± SD. ** P < 0.01 vs. CTL-NC; # P < 0.05 vs. CKO-NC; # # P < 0.01 vs. CKO-NC; # ### P < 0.01 vs. CTL-STZ. (D) Representative images of PAS and MASSON staining of the glomeruli in mice at 12 and 16 weeks. Scale bar, 20 μm ( n = 8). (E) Representative images of HE staining of the kidney in mice at 12 and 16 weeks. Scale bar, 50 μm ( n = 8). (F) Representative IF images of the expression of SD proteins p-cadherin and <t>synaptopodin</t> in glomeruli at 12 and 16 weeks. Scale bar, 20 μm ( n = 8). (G) The expression of a mesenchymal marker desmin was detected in renal tissues from mice at 12 and 16 weeks by western blot analysis. P values were determined by one-way ANOVA and data are presented as mean ± SD. **** P < 0.01; # P < 0.05; ## P < 0.05 ( n = 8). (H) Representative images of TEM in podocyte ultrastructural alterations in mice. FPs effacement (blue arrow), ER dilation (red arrow) and GBM thickness were observed in podocytes in STZ-induced CTL diabetic mice, and were attenuated in TLR4 CKO diabetic mice at 16 weeks. Scale bar, 1 μm ( n = 8). (I) Determination of proteinuria in mice. Data are presented as mean ± SD. ** P < 0.01 vs. CTL-NC; # P < 0.05 vs. CKO-NC; # # P < 0.01 vs. CKO-NC; # ## P < 0.05 vs. CTL-STZ; # ### P < 0.01 vs. CTL-STZ.
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    Image Search Results


    Podocyte-specific TLR4 deletion mitigated STZ-induced DKD, podocyte injury, and proteinuria in mice. (A) Schematic description of the strategy for generating STZ-induced diabetic mouse model in NPHS2 -Cre; TLR4 fl/fl ( TLR4 CKO) mice and in TLR4 fl/fl mice (CTL). (B) Monthly monitoring of plasma glucose levels in mice. Data are presented as mean ± SD. ** P < 0.01 vs. CTL-NC; ## P < 0.01 vs. CKO-NC. (C) Monthly monitoring of body weight in mice. Data are presented as mean ± SD. ** P < 0.01 vs. CTL-NC; # P < 0.05 vs. CKO-NC; # # P < 0.01 vs. CKO-NC; # ### P < 0.01 vs. CTL-STZ. (D) Representative images of PAS and MASSON staining of the glomeruli in mice at 12 and 16 weeks. Scale bar, 20 μm ( n = 8). (E) Representative images of HE staining of the kidney in mice at 12 and 16 weeks. Scale bar, 50 μm ( n = 8). (F) Representative IF images of the expression of SD proteins p-cadherin and synaptopodin in glomeruli at 12 and 16 weeks. Scale bar, 20 μm ( n = 8). (G) The expression of a mesenchymal marker desmin was detected in renal tissues from mice at 12 and 16 weeks by western blot analysis. P values were determined by one-way ANOVA and data are presented as mean ± SD. **** P < 0.01; # P < 0.05; ## P < 0.05 ( n = 8). (H) Representative images of TEM in podocyte ultrastructural alterations in mice. FPs effacement (blue arrow), ER dilation (red arrow) and GBM thickness were observed in podocytes in STZ-induced CTL diabetic mice, and were attenuated in TLR4 CKO diabetic mice at 16 weeks. Scale bar, 1 μm ( n = 8). (I) Determination of proteinuria in mice. Data are presented as mean ± SD. ** P < 0.01 vs. CTL-NC; # P < 0.05 vs. CKO-NC; # # P < 0.01 vs. CKO-NC; # ## P < 0.05 vs. CTL-STZ; # ### P < 0.01 vs. CTL-STZ.

    Journal: Journal of Advanced Research

    Article Title: Podocyte TLR4 deletion alleviates diabetic kidney disease through prohibiting PKCδ/SHP-1-dependent ER stress and relieving podocyte damage and inflammation

    doi: 10.1016/j.jare.2025.07.013

    Figure Lengend Snippet: Podocyte-specific TLR4 deletion mitigated STZ-induced DKD, podocyte injury, and proteinuria in mice. (A) Schematic description of the strategy for generating STZ-induced diabetic mouse model in NPHS2 -Cre; TLR4 fl/fl ( TLR4 CKO) mice and in TLR4 fl/fl mice (CTL). (B) Monthly monitoring of plasma glucose levels in mice. Data are presented as mean ± SD. ** P < 0.01 vs. CTL-NC; ## P < 0.01 vs. CKO-NC. (C) Monthly monitoring of body weight in mice. Data are presented as mean ± SD. ** P < 0.01 vs. CTL-NC; # P < 0.05 vs. CKO-NC; # # P < 0.01 vs. CKO-NC; # ### P < 0.01 vs. CTL-STZ. (D) Representative images of PAS and MASSON staining of the glomeruli in mice at 12 and 16 weeks. Scale bar, 20 μm ( n = 8). (E) Representative images of HE staining of the kidney in mice at 12 and 16 weeks. Scale bar, 50 μm ( n = 8). (F) Representative IF images of the expression of SD proteins p-cadherin and synaptopodin in glomeruli at 12 and 16 weeks. Scale bar, 20 μm ( n = 8). (G) The expression of a mesenchymal marker desmin was detected in renal tissues from mice at 12 and 16 weeks by western blot analysis. P values were determined by one-way ANOVA and data are presented as mean ± SD. **** P < 0.01; # P < 0.05; ## P < 0.05 ( n = 8). (H) Representative images of TEM in podocyte ultrastructural alterations in mice. FPs effacement (blue arrow), ER dilation (red arrow) and GBM thickness were observed in podocytes in STZ-induced CTL diabetic mice, and were attenuated in TLR4 CKO diabetic mice at 16 weeks. Scale bar, 1 μm ( n = 8). (I) Determination of proteinuria in mice. Data are presented as mean ± SD. ** P < 0.01 vs. CTL-NC; # P < 0.05 vs. CKO-NC; # # P < 0.01 vs. CKO-NC; # ## P < 0.05 vs. CTL-STZ; # ### P < 0.01 vs. CTL-STZ.

    Article Snippet: The following primary antibodies were used: f4/80 (29414–1-AP), desmin (16520–1-AP), MCP-1 (26161–1-AP), podocin (20384–1-AP), synaptopodin (21064–1-AP), TLR4 (66350–1-Ig), β-actin (66009–1-Ig), PKCδ(14188–1-AP), ATF6 (24169–1-AP), p-cadherin (13773–1-AP) were from Proteintech (IL, USA); phospho-PKCδ (Y311) (ab76181), PKCdelta (ab182126), SHP-1 (ab227503), HA (ab9110) were from Abcam (MA, USA); BIP (M010300), MyD88 (P012343) were from Epizyme, Shanghai, China); and ATF4 (11815), sXBP-1 (40435) were from Cell Signaling Technology (MA, USA).

    Techniques: Clinical Proteomics, Staining, Expressing, Marker, Western Blot

    Activation of TLR4/MyD88/PKCδ/SHP-1 signaling cascade was involved in hyperglycemic podocyte damage in vivo and in vitro . (A) The protein levels of TLR4 were determined in renal tissues from mice at 12 and 16 weeks by western blot analysis. P values were determined by one-way ANOVA and data are presented as mean ± SD. **** P < 0.01; # P < 0.05 ( n = 8). (B) Representative western blot of the expression of MyD88, p-PKCδ (Y311), PKCδ and SHP-1 in renal cortex from mice at 12 and 16 weeks. P values were determined by one-way ANOVA and data are presented as mean ± SD. **** P < 0.01; # P < 0.05; ## P < 0.01; ### P < 0.01 ( n = 8). (C) Representative double IF staining of glomerular SHP-1 and synaptopodin expression in mice at 16 weeks. Scale bar, 20 μm ( n = 8). (D) The expression of TLR4, MyD88, p-PKCδ, PKCδ and SHP-1 in cultured mouse podocytes from treatments of LG or HG for 48 h with or without transfection of TLR4, MyD88 or scrambled shRNAs. P values were determined by one-way ANOVA and data are presented as mean ± SD. **** P < 0.01; ## P < 0.01; ### P < 0.01; #### P < 0.01 ( n = 3 independent experiments). (E) IF staining of podocyte proteins p-cadherin and synaptopodin after LG or HG stimulation for 48 h with or without transfection of TLR4, PKCδ, MCP-1 or scrambled shRNAs, or pretreatment of 4-PBA. Magnification 400 × . ( n = 3 independent experiments). (F) Determination of SD protein podocin and injury marker desmin in podocytes treated with LG or HG for 48 h with or without transfection of TLR4, MyD88 or scrambled shRNAs. P values were determined by one-way ANOVA and data are presented as mean ± SD. **** P < 0.01; ### P < 0.01; #### P < 0.01 ( n = 3 independent experiments).

    Journal: Journal of Advanced Research

    Article Title: Podocyte TLR4 deletion alleviates diabetic kidney disease through prohibiting PKCδ/SHP-1-dependent ER stress and relieving podocyte damage and inflammation

    doi: 10.1016/j.jare.2025.07.013

    Figure Lengend Snippet: Activation of TLR4/MyD88/PKCδ/SHP-1 signaling cascade was involved in hyperglycemic podocyte damage in vivo and in vitro . (A) The protein levels of TLR4 were determined in renal tissues from mice at 12 and 16 weeks by western blot analysis. P values were determined by one-way ANOVA and data are presented as mean ± SD. **** P < 0.01; # P < 0.05 ( n = 8). (B) Representative western blot of the expression of MyD88, p-PKCδ (Y311), PKCδ and SHP-1 in renal cortex from mice at 12 and 16 weeks. P values were determined by one-way ANOVA and data are presented as mean ± SD. **** P < 0.01; # P < 0.05; ## P < 0.01; ### P < 0.01 ( n = 8). (C) Representative double IF staining of glomerular SHP-1 and synaptopodin expression in mice at 16 weeks. Scale bar, 20 μm ( n = 8). (D) The expression of TLR4, MyD88, p-PKCδ, PKCδ and SHP-1 in cultured mouse podocytes from treatments of LG or HG for 48 h with or without transfection of TLR4, MyD88 or scrambled shRNAs. P values were determined by one-way ANOVA and data are presented as mean ± SD. **** P < 0.01; ## P < 0.01; ### P < 0.01; #### P < 0.01 ( n = 3 independent experiments). (E) IF staining of podocyte proteins p-cadherin and synaptopodin after LG or HG stimulation for 48 h with or without transfection of TLR4, PKCδ, MCP-1 or scrambled shRNAs, or pretreatment of 4-PBA. Magnification 400 × . ( n = 3 independent experiments). (F) Determination of SD protein podocin and injury marker desmin in podocytes treated with LG or HG for 48 h with or without transfection of TLR4, MyD88 or scrambled shRNAs. P values were determined by one-way ANOVA and data are presented as mean ± SD. **** P < 0.01; ### P < 0.01; #### P < 0.01 ( n = 3 independent experiments).

    Article Snippet: The following primary antibodies were used: f4/80 (29414–1-AP), desmin (16520–1-AP), MCP-1 (26161–1-AP), podocin (20384–1-AP), synaptopodin (21064–1-AP), TLR4 (66350–1-Ig), β-actin (66009–1-Ig), PKCδ(14188–1-AP), ATF6 (24169–1-AP), p-cadherin (13773–1-AP) were from Proteintech (IL, USA); phospho-PKCδ (Y311) (ab76181), PKCdelta (ab182126), SHP-1 (ab227503), HA (ab9110) were from Abcam (MA, USA); BIP (M010300), MyD88 (P012343) were from Epizyme, Shanghai, China); and ATF4 (11815), sXBP-1 (40435) were from Cell Signaling Technology (MA, USA).

    Techniques: Activation Assay, In Vivo, In Vitro, Western Blot, Expressing, Staining, Cell Culture, Transfection, Marker

    TLR4/MyD88/PKCδ/SHP-1 signaling cascade induced podocyte damage through regulating ER stress in DKD and podocyte damage. (A) The protein levels of ER stress markers BIP, sXBP-1, ATF4 and ATF6 were determined in renal tissues from mice at 12 and 16 weeks by western blot. P values were determined by one-way ANOVA and data are presented as mean ± SD. **** P < 0.01; # P < 0.05; ## P < 0.01; ### P < 0.01 ( n = 8). (B) Representative double IF staining of the expression ATF4 and podocyte marker synaptopodin in mice at 16 weeks. Scale bar, 20 μm ( n = 8). (C) Representative western blot to assess the levels of ER stress markers BIP, sXBP-1, ATF4 and ATF6 in cultured mouse podocytes from treatments of LG or HG for 48 h with or without transfection of TLR4, MyD88 or scrambled shRNAs. P values were determined by one-way ANOVA and data are presented as mean ± SD. **** P < 0.01; ### P < 0.01; ### P < 0.01 ( n = 3 independent experiments). (D) ER morphological alterations and ER stress illustrated by ER-tracker staining in podocytes treated with LG or HG for 48 h with or without transfection of TLR4, PKCδ, or MCP-1 shRNAs, or pretreatment of 4-PBA. Scale bar, 10 μm. P values were determined by one-way ANOVA and data are presented as mean ± SD. **** P < 0.01; ## P < 0.01; # P < 0.05 ( n = 3 independent experiments). (E) Representative western blot to detect the expression of ER stress proteins BIP, sXBP-1, ATF4 and ATF6 in cultured podocytes after HG stimulation for 48 h with or without transfection of PKCδ, SHP-1 or scrambled shRNAs, or pretreatment of 4-PBA. P values were determined by one-way ANOVA and data are presented as mean ± SD. ** P < 0.01; *** P < 0.01; **** P < 0.01; ## P < 0.01; ### P < 0.01; #### P < 0.01 ( n = 3 independent experiments).

    Journal: Journal of Advanced Research

    Article Title: Podocyte TLR4 deletion alleviates diabetic kidney disease through prohibiting PKCδ/SHP-1-dependent ER stress and relieving podocyte damage and inflammation

    doi: 10.1016/j.jare.2025.07.013

    Figure Lengend Snippet: TLR4/MyD88/PKCδ/SHP-1 signaling cascade induced podocyte damage through regulating ER stress in DKD and podocyte damage. (A) The protein levels of ER stress markers BIP, sXBP-1, ATF4 and ATF6 were determined in renal tissues from mice at 12 and 16 weeks by western blot. P values were determined by one-way ANOVA and data are presented as mean ± SD. **** P < 0.01; # P < 0.05; ## P < 0.01; ### P < 0.01 ( n = 8). (B) Representative double IF staining of the expression ATF4 and podocyte marker synaptopodin in mice at 16 weeks. Scale bar, 20 μm ( n = 8). (C) Representative western blot to assess the levels of ER stress markers BIP, sXBP-1, ATF4 and ATF6 in cultured mouse podocytes from treatments of LG or HG for 48 h with or without transfection of TLR4, MyD88 or scrambled shRNAs. P values were determined by one-way ANOVA and data are presented as mean ± SD. **** P < 0.01; ### P < 0.01; ### P < 0.01 ( n = 3 independent experiments). (D) ER morphological alterations and ER stress illustrated by ER-tracker staining in podocytes treated with LG or HG for 48 h with or without transfection of TLR4, PKCδ, or MCP-1 shRNAs, or pretreatment of 4-PBA. Scale bar, 10 μm. P values were determined by one-way ANOVA and data are presented as mean ± SD. **** P < 0.01; ## P < 0.01; # P < 0.05 ( n = 3 independent experiments). (E) Representative western blot to detect the expression of ER stress proteins BIP, sXBP-1, ATF4 and ATF6 in cultured podocytes after HG stimulation for 48 h with or without transfection of PKCδ, SHP-1 or scrambled shRNAs, or pretreatment of 4-PBA. P values were determined by one-way ANOVA and data are presented as mean ± SD. ** P < 0.01; *** P < 0.01; **** P < 0.01; ## P < 0.01; ### P < 0.01; #### P < 0.01 ( n = 3 independent experiments).

    Article Snippet: The following primary antibodies were used: f4/80 (29414–1-AP), desmin (16520–1-AP), MCP-1 (26161–1-AP), podocin (20384–1-AP), synaptopodin (21064–1-AP), TLR4 (66350–1-Ig), β-actin (66009–1-Ig), PKCδ(14188–1-AP), ATF6 (24169–1-AP), p-cadherin (13773–1-AP) were from Proteintech (IL, USA); phospho-PKCδ (Y311) (ab76181), PKCdelta (ab182126), SHP-1 (ab227503), HA (ab9110) were from Abcam (MA, USA); BIP (M010300), MyD88 (P012343) were from Epizyme, Shanghai, China); and ATF4 (11815), sXBP-1 (40435) were from Cell Signaling Technology (MA, USA).

    Techniques: Western Blot, Staining, Expressing, Marker, Cell Culture, Transfection