nephrin  (Bioss)

Journal: Frontiers in Pharmacology

Article Title: Hyperoside ameliorates lupus nephritis by suppressing AKT1-mediated PANoptosis in podocytes: integrating network pharmacology and experimental validation

doi: 10.3389/fphar.2025.1726254

Figure Lengend Snippet: PANoptosis contributes to renal pathology in lupus nephritis. (A) Comparison of 24-h urinary protein levels between MRL/MpJ and MRL/lpr mice (n = 6). (B) Representative renal histology evaluated by H&E staining (n = 6). (C) TUNEL staining showing cell death in glomeruli (n = 6). (D) IHC analysis of podocyte markers (Podocalyxin, Nephrin, Synaptopodin, Podocin) in renal tissues. (E–G) Immunofluorescence staining of PANoptosis-related markers (NLRP3, RIPK3, Caspase-9) in kidney sections. (H) WB analysis of PANoptosis executor proteins (caspase-3, caspase-1, p-MLKL) in renal tissues, with bar graphs showing quantitative results relative to GAPDH (n = 3). (I) IF triple-staining of caspase-3, caspase-1, and p-MLKL in mouse kidney tissues. (J) Viability of MPC-5 cells treated with ICs at different concentrations and time points, measured by CCK-8 assay. (K) Flow cytometric analysis of cell death in MPC5 cells pretreated with VX-765 (10 μM), Nec-1 (20 μM), or Z-VAD (5 μM), followed by IC stimulation for 24 h. Bar graph shows the percentage of dead cells (n = 3). Data are presented as mean ± SD. * p < 0.05, ** p < 0.01, ns: not significant.

Article Snippet: Sequential sections (4 μm) were used for the following staining and analyses: Hematoxylin and eosin (H&E) staining was performed to evaluate pathological changes in renal tissues; immunohistochemistry (IHC) was conducted to examine the expression of synaptopodin (21064-1-AP, Proteintech, USA), nephrin (bs-10233R, Bioss, China), podocalyxin (bs-1345R, Bioss, China), and podocin (bs-6597R, Bioss, China); immunofluorescence (IF) staining was used to detect NLRP3 (DF7438, Affinity, UK), RIPK3 (bs-3551R, Bioss, China), and Caspase-9 (bs-0049R, Bioss, China) expression; and a TUNEL assay kit (E-CK-A320, Elabscience, China) was employed to assess cell death.

Techniques: Comparison, Staining, TUNEL Assay, Immunofluorescence, CCK-8 Assay

Journal: Frontiers in Pharmacology

Article Title: Hyperoside ameliorates lupus nephritis by suppressing AKT1-mediated PANoptosis in podocytes: integrating network pharmacology and experimental validation

doi: 10.3389/fphar.2025.1726254

Figure Lengend Snippet: AKT1 mediates podocyte PANoptosis and LN progression. (A–E) MRL/lpr mice were injected via tail vein with KD-NC or KD-AKT1 AAV and maintained for 6 weeks (n = 6). (A,B) Western blot analysis of AKT1 and PANoptosis markers (p-MLKL, Caspase-1, Caspase-3) in renal tissues (n = 3). (C) Renal pathology assessed by H&E staining. (D) Immunohistochemical staining of podocyte-associated proteins (Podocalyxin, Nephrin, Synaptopodin, Podocin). (E) Comparison of 24 h urinary protein levels between the two groups. (F–P) MPC-5 cells were transfected with Si-NC, Si-AKT1, OE-NC, or OE-AKT1, followed by IC treatment for 24 h. (F) Percentage of dead cells measured by flow cytometry (n = 6). (G) Representative flow cytometry plots showing apoptosis. (H–J) mRNA expression levels of PANoptosis-related genes (NLRP3, RIPK3, Caspase-9) detected by qPCR (n = 6). (K–P) Western blot analysis of PANoptosis executor proteins (p-MLKL, Caspase-1, Caspase-3, GSDMD-N, Caspase-8) in MPC5 cells (n = 3). Data are presented as mean ± SD. * p < 0.05, ** p < 0.01, ns: not significant.

Article Snippet: Sequential sections (4 μm) were used for the following staining and analyses: Hematoxylin and eosin (H&E) staining was performed to evaluate pathological changes in renal tissues; immunohistochemistry (IHC) was conducted to examine the expression of synaptopodin (21064-1-AP, Proteintech, USA), nephrin (bs-10233R, Bioss, China), podocalyxin (bs-1345R, Bioss, China), and podocin (bs-6597R, Bioss, China); immunofluorescence (IF) staining was used to detect NLRP3 (DF7438, Affinity, UK), RIPK3 (bs-3551R, Bioss, China), and Caspase-9 (bs-0049R, Bioss, China) expression; and a TUNEL assay kit (E-CK-A320, Elabscience, China) was employed to assess cell death.

Techniques: Injection, Western Blot, Staining, Immunohistochemical staining, Comparison, Transfection, Flow Cytometry, Expressing

Journal: APL Bioengineering

Article Title: Geometrically engineered organoid units and their assembly for pre-construction of organ structures

doi: 10.1063/5.0222866

Figure Lengend Snippet: Construction and assembly of human kidney glomerular tissue-like organoid units. (a) Fluorescence-stained frozen sections of tri-cultured spherical organoid units ( n = 3). Each marker protein is stained on three cell types: NHMCs (CD90.1), HUVECs (CD31), and HRGEpCs (Nephrin). Scale bar, 50 μ m. (b) Changes over culture duration in co-localization of marker proteins (mean ± SD, n = 3; 8; or 9 images). (c) Overall view of fluorescence-stained frozen sections of tri-cultured spherical organoid units ( n = 3). Scale bar, 500 μ m. (d) Representative fluorescence-stained images of cellular dynamics around the bonding area between the tri-cultured cubic organoid units ( n = 3, upper row : overall view. Scale bar, 1 mm; lower 5 rows : magnified view. Scale bar, 500 μ m).

Article Snippet: The sheep polyclonal anti-nephrin antibody (Cat# AF4269) was purchased from R&D Systems (Minneapolis, MN, USA).

Techniques: Fluorescence, Staining, Cell Culture, Marker

Journal: APL Bioengineering

Article Title: Geometrically engineered organoid units and their assembly for pre-construction of organ structures

doi: 10.1063/5.0222866

Figure Lengend Snippet: Assembly of organoid units with different geometries. (a) Overview of how to assemble the organoid units with different geometries. (b) Time-sequenced brightfield images of the assembled MDA-MB-231 organoid units ( n = 3). Scale bar, 1 mm. (c) Live fluorescence images of the assembled MDA-MB-231 organoid units, expressing GFP in the cytoplasm ( n = 3). Upper row : wide-field fluorescence images (raw data), lower row : images processed by the THUNDER imaging system with instant computational clearing (ICC) and extended depth of field (EDF). Scale bars, 1 mm. (d) Representative fluorescence-stained images of the assembled human-kidney glomerular tissue-like organoid units ( n = 3). Each marker protein is stained on three cell types: NHMCs (CD90.1), HUVECs (CD31), and HRGEpCs (Nephrin). Upper 5 rows : wide-field fluorescence images (raw data), lower row : images processed by the THUNDER imaging system with instant computational clearing (ICC) and extended depth of field (EDF). Scale bar, 1 mm.

Article Snippet: The sheep polyclonal anti-nephrin antibody (Cat# AF4269) was purchased from R&D Systems (Minneapolis, MN, USA).

Techniques: Fluorescence, Expressing, Imaging, Staining, Marker