pas glycogen staining kit  (Beyotime)

Journal: eLife

Article Title: Glycogen engineering improves the starvation resistance of mesenchymal stem cells and their therapeutic efficacy in pulmonary fibrosis

doi: 10.7554/eLife.106023

Figure Lengend Snippet: ( A ) Essential enzymes of glycogen synthesis. ( B ) Tests of essential enzyme combination strategies in HEK293T cells by transient transfection. The glycogen content of each group was measured (N=3, mean ± SD). ( C ) Periodic acid-Schiff (PAS) staining of cells expressing GYSmut and glycogen synthase (GYS)-UDP-glucose pyrophosphorylase (UGP) revealed significant accumulation of glycogen granules.Scale bar, 50 μm.

Article Snippet: A PAS staining kit (Beyotime, C0142S) was used to stain glycogen granules.

Techniques: Transfection, Staining, Expressing

Journal: eLife

Article Title: Glycogen engineering improves the starvation resistance of mesenchymal stem cells and their therapeutic efficacy in pulmonary fibrosis

doi: 10.7554/eLife.106023

Figure Lengend Snippet: ( A ) Glycogen content of GYSmut MSCs (N=3, mean ± SD). ( B ) Periodic acid-Schiff (PAS) staining of GYSmut MSCs, showing glycogen granules (red). Scale bar, 50 μm. ( C ) Survival of engineered MSCs under Dulbecco’s phosphate-buffered saline (DPBS) (starvation) treatment in vitro (N=3, mean ± SD). ( D ) Residual glycogen content of GYSmut MSCs after DPBS (starvation) treatment for 48 hr (N=3, mean ± SD). ( E ) Viability of GYSmut MSCs according to the CCK8 assay (N=8, mean ± SD). ( F, G ) Adipogenic differentiation potential of GYSmut MSCs, assessed by Oil Red O staining and qPCR detection of Lpl expression (N=3, mean ± SD, unpaired t -test p-value<0.0001). Scale bar, 100 μm. ( H ) Gene ontology (GO) enrichment analysis of differentially expressed genes (DEGs) between GYSmut MSCs and the GFP control. ( I ) KEGG analysis of DEGs. ( J ) Gene set enrichment analysis (GSEA) of the DEGs.

Article Snippet: A PAS staining kit (Beyotime, C0142S) was used to stain glycogen granules.

Techniques: Staining, Saline, In Vitro, CCK-8 Assay, Expressing, Control

Journal: eLife

Article Title: Glycogen engineering improves the starvation resistance of mesenchymal stem cells and their therapeutic efficacy in pulmonary fibrosis

doi: 10.7554/eLife.106023

Figure Lengend Snippet: ( A ) Cell volume change of engineered MSCs, assessed through flow cytometry. ( B ) Periodic acid-Schiff (PAS) and hematoxylin staining of GYSmut MSCs. Glycogen (red) is distributed in cell nucleus (blue) and cytoplasm. ( C ) Glycogen distribution of GYSmut and glycogen synthase (GYS)-glycogenin (GYG) MSCs. Scale bar, 50 μm. ( D ) Starvation resistance test under hypoxia.(N=5, mean ± SD). ( E ) Impacts of glycogen engineering on transcriptome of MSCs.

Article Snippet: A PAS staining kit (Beyotime, C0142S) was used to stain glycogen granules.

Techniques: Flow Cytometry, Staining

Journal: The FASEB Journal

Article Title: SMYD2 Promotes Renal Tubular Cell Apoptosis and Chronic Kidney Disease Following Cisplatin Nephrotoxicity

doi: 10.1096/fj.202402703R

Figure Lengend Snippet: AZ505 mitigates renal injury and suppresses the upregulation of ECM proteins in the renal tissue of mice with cisplatin‐induced CKD. (A, B) Serum blood urea nitrogen (BUN) and serum creatinine (Scr) levels in AZ505‐treated and untreated cisplatin‐induced CKD mice ( n = 5). (C) Hematoxylin and eosin (H&E), Masson's trichrome, and periodic acid‐Schiff (PAS) staining of renal tissue sections from AZ505‐treated and untreated cisplatin‐induced CKD mice. Scale bar: 10 μm. (D, E) Western blot analysis of ECM protein expression in renal tissues from AZ505‐treated and untreated cisplatin‐induced CKD mice ( n = 3). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: DMEM/F12 medium, fetal bovine serum (FBS) were purchased from Invitrogen Gibco, USA; tryptic digest, penicillin–streptomycin mixture were purchased from Biological Industries, Israel; Cis‐platin (CIS), SMYD2 specific inhibitor AZ505, NF‐κB specific inhibitor BAY11‐7085, Streptavidin‐FITC were purchased from APExBIO, USA; Smart‐ECL Enhanced Luminescence Kit was purchased from Changzhou Tiandi Renhe Bio‐technology Co. Ltd.; Hematoxylin and Eosin (HE) kit, MASSON Trichrome Staining Improvement Kit, Glycogen Staining (Periodic Acid Schiff (PAS)) kit, BCA Protein Concentration Measurement Kit, CCK‐8 (Cell Counting Kit‐8, CCK‐8) kit, Streptavidin‐FITC were purchased from APExBIO, USA. high‐efficiency RIPA (Radio Immuno Precipitation Assay, RIPA) lysate, anti‐fluorescence attenuation sealer containing 4′,6‐diamidino‐2‐phenylindole (DAPI), Bromphenol Blue, DL‐Dithiothreitol (DTT), NP‐40 LYSIS BUFFER lysate, and protease inhibitor (Phenylmethanesulfonyl fluoride (PMSF)) were purchased from Beijing Soleilbao Technology Co. TB GreeTM Premix Ex TaqTM II (TLi RNaseH Plus) was purchased from Takara Bio, Japan; Hifair III 1st Strand cDNA Synthesis SuperMix for qPCR (gDNA digester plus) was purchased from Yi Sheng Biotechnology (Shanghai); TUNEL kit (ElabscienceOne‐step TUNEL In Situ Apoptosis Kit) was purchased from Wuhan Elabscience Biotechnology Co. LTD; Goat anti‐Mouse IgG (H + L), CY3 AffiniPure was purchased from Wuhan Pramerica Biotechnology Co. biom; LTL, Biotinylated was purchased from Vector Laboratories Inc. in the USA; Aquaporin 1 (AQP1) (sc‐25 287), Protein A/G PLUS‐Agarose was purchased from SANTA CRUZ biotechnology Co. SANTA CRUZ biotechnology Inc.

Techniques: Staining, Western Blot, Expressing

Journal: The FASEB Journal

Article Title: SMYD2 Promotes Renal Tubular Cell Apoptosis and Chronic Kidney Disease Following Cisplatin Nephrotoxicity

doi: 10.1096/fj.202402703R

Figure Lengend Snippet: Smyd2 tecKO Mitigates kidney injury in cisplatin‐induced CKD mice and suppresses ECM protein upregulation. (A–C) Immunofluorescence and Western blot analysis of SMYD2 expression and localization in renal tissues of Smyd2 tecKO mice, with β‐Actin as a loading control (scale bar: 50 μm; N = 3). (D) Schematic of cisplatin‐induced CKD model in Smyd2 tecKO mice. (E–G) Blood urea nitrogen (BUN), serum creatinine (Scr), and proteinuria levels in cisplatin‐treated Smyd2 tecKO mice and Smyd2 fl/fl mice ( n = 5). (H) H&E, Masson, and PAS staining of kidney sections. Scale bar: 10 μm. (I, J) Western blot analysis of renal ECM proteins (α‐SMA, fibronectin, collagen I) in cisplatin‐treated groups ( n = 3). * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: DMEM/F12 medium, fetal bovine serum (FBS) were purchased from Invitrogen Gibco, USA; tryptic digest, penicillin–streptomycin mixture were purchased from Biological Industries, Israel; Cis‐platin (CIS), SMYD2 specific inhibitor AZ505, NF‐κB specific inhibitor BAY11‐7085, Streptavidin‐FITC were purchased from APExBIO, USA; Smart‐ECL Enhanced Luminescence Kit was purchased from Changzhou Tiandi Renhe Bio‐technology Co. Ltd.; Hematoxylin and Eosin (HE) kit, MASSON Trichrome Staining Improvement Kit, Glycogen Staining (Periodic Acid Schiff (PAS)) kit, BCA Protein Concentration Measurement Kit, CCK‐8 (Cell Counting Kit‐8, CCK‐8) kit, Streptavidin‐FITC were purchased from APExBIO, USA. high‐efficiency RIPA (Radio Immuno Precipitation Assay, RIPA) lysate, anti‐fluorescence attenuation sealer containing 4′,6‐diamidino‐2‐phenylindole (DAPI), Bromphenol Blue, DL‐Dithiothreitol (DTT), NP‐40 LYSIS BUFFER lysate, and protease inhibitor (Phenylmethanesulfonyl fluoride (PMSF)) were purchased from Beijing Soleilbao Technology Co. TB GreeTM Premix Ex TaqTM II (TLi RNaseH Plus) was purchased from Takara Bio, Japan; Hifair III 1st Strand cDNA Synthesis SuperMix for qPCR (gDNA digester plus) was purchased from Yi Sheng Biotechnology (Shanghai); TUNEL kit (ElabscienceOne‐step TUNEL In Situ Apoptosis Kit) was purchased from Wuhan Elabscience Biotechnology Co. LTD; Goat anti‐Mouse IgG (H + L), CY3 AffiniPure was purchased from Wuhan Pramerica Biotechnology Co. biom; LTL, Biotinylated was purchased from Vector Laboratories Inc. in the USA; Aquaporin 1 (AQP1) (sc‐25 287), Protein A/G PLUS‐Agarose was purchased from SANTA CRUZ biotechnology Co. SANTA CRUZ biotechnology Inc.

Techniques: Immunofluorescence, Western Blot, Expressing, Control, Staining

Journal: Frontiers in Microbiology

Article Title: Clostridium butyricum ameliorates indomethacin-induced enteropathy by promoting MUC2 secretion via suppressing the Notch pathway

doi: 10.3389/fmicb.2025.1509876

Figure Lengend Snippet: Clostridium butyricum (CB) increased the number of goblet cells and MUC2 secretion in the small intestine. (A,C) Goblet cells in the intestine were assessed by PAS staining, and the number of positive cells in each intestinal villus was calculated (scale bar = 200 μm, n = 6). (B,D) MUC2 expression in the intestine was detected by immunohistochemistry, and the number of positive cells in each intestinal villus was calculated (scale bar = 200 μm, n = 3). (E) Relative mRNA expression of MUC2 in intestinal tissues, as analyzed via qRT-PCR ( n = 6). Data were presented as mean ± standard deviation. * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: For measurement of the number of goblet cells, paraffin sections were subjected to PAS staining in accordance with the instructions of the glycogen PAS staining kit (Solarbio, China).

Techniques: Staining, Expressing, Immunohistochemistry, Quantitative RT-PCR, Standard Deviation