Journal: Gut and Liver

Article Title: The Use of Foxa2-Overexpressing Adipose Tissue-Derived Stem Cells in a Scaffold System Attenuates Acute Liver Injury

doi: 10.5009/gnl18235

Figure Lengend Snippet: Hepatic differentiation ability in two-dimensional (2D) culture. (A) Gene expression of hepatic differentiation markers (AFP, ALB, and CK18) in transfected ADSCs after 2 weeks measured by qRT-PCR. (B) Amount of cytoplasmic glycogen after the overexpression of various gene in 2D culture measured by periodic acid-Schiff (PAS) staining (PAS staining: blue, nuclei; purple, glycogen in cytoplasm; magnification: ×400). The arrows indicate cytosol. AFP, alpha fetoprotein, ALB, albumin; CK18, cytokeratin 18; ADSCs, adipose tissue-derived stem cells; qRT-PCR, real-time quantitative polymerase chain reaction; Foxa2, forkhead box protein. *p<0.05.

Article Snippet: For liver function evaluation, periodic acid-Schiff staining (PAS Staining Kit, 1.01646.0001; Merck Millipore) was performed to analyze glycogen storage capacity.

Techniques: Gene Expression, Transfection, Quantitative RT-PCR, Over Expression, Staining, Derivative Assay, Real-time Polymerase Chain Reaction

Journal: Gut and Liver

Article Title: The Use of Foxa2-Overexpressing Adipose Tissue-Derived Stem Cells in a Scaffold System Attenuates Acute Liver Injury

doi: 10.5009/gnl18235

Figure Lengend Snippet: Comparison of hepatic differentiation ability in three-dimensional (3D) culture using a scaffold. (A) qRT-PCR expression of hepatic differentiation markers (AFP, CK18, CD26, and CX32) in Foxa2-transfected ADSCs after 2 weeks in 3D culture. (B) H&E staining, periodic acid-Schiff (PAS) and cytochrome P450 immunofluorescence in Foxa2-overexpressing ADSCs in 3D culture (a and b: H&E staining, blue - nuclei, pink - cytosol, ×400; c and d: PAS staining, blue - nuclei, purple - glycogen in cytoplasm, ×1,000; e and f: blue - nuclei, green - cytochrome 450 in cytosol, ×1,000). (C) Quantification of cytochrome P450 intensity and glucose expression. qRT-PCR, real-time quantitative polymerase chain reaction; AFP, alpha fetoprotein; CK18, cytokeratin 18; CD26, dipeptidyl peptidase 4; CX32, connexin 32; ADSCs, adipose tissue-derived stem cells; Foxa2, forkhead box protein. *p<0.05.

Article Snippet: For liver function evaluation, periodic acid-Schiff staining (PAS Staining Kit, 1.01646.0001; Merck Millipore) was performed to analyze glycogen storage capacity.

Techniques: Comparison, Quantitative RT-PCR, Expressing, Transfection, Staining, Immunofluorescence, Real-time Polymerase Chain Reaction, Derivative Assay

Journal: Gut and Liver

Article Title: The Use of Foxa2-Overexpressing Adipose Tissue-Derived Stem Cells in a Scaffold System Attenuates Acute Liver Injury

doi: 10.5009/gnl18235

Figure Lengend Snippet: Comparison of hepatocyte differentiation in a scaffold during in vivo culture. (A) In vivo culture design of scaffold preparation and implantation into nude mice. (B) Scanning electron microscopy of ADSCs in the PLGA scaffold. (C) qRT-PCR expression of hepatic differentiation markers (AFP, CK18, CD26 and CX32) in Foxa2-transfected ADSCs after 2 weeks in nude mice. (D) H&E staining, periodic acid-Schiff (PAS) staining and immunofluorescent staining for cytochrome P450 activity after Foxa2 gene overexpression in in vivo culture (a and b: H&E staining, blue - nuclei, pink - cytosol, ×400; c and d: PAS staining, blue - nuclei, purple - glycogen in cytoplasm, ×1,000; e and f: blue - nuclei, green - cytochrome 450 in cytosol, ×1,000). (E) Quantification of cytochrome P450 intensity and glucose expression. ADSCs, adipose tissue-derived stem cells; PLGA, poly(lactic-co-glycolic acid); qRT-PCR, real-time quantitative polymerase chain reaction; AFP, alpha fetoprotein; CK18, cytokeratin 18; CD26, dipeptidyl peptidase 4; CX32, connexin 32; Foxa2, forkhead box protein. *p<0.05.

Article Snippet: For liver function evaluation, periodic acid-Schiff staining (PAS Staining Kit, 1.01646.0001; Merck Millipore) was performed to analyze glycogen storage capacity.

Techniques: Comparison, In Vivo, Electron Microscopy, Quantitative RT-PCR, Expressing, Transfection, Staining, Activity Assay, Over Expression, Derivative Assay, Real-time Polymerase Chain Reaction

Journal: Gut and Liver

Article Title: The Use of Foxa2-Overexpressing Adipose Tissue-Derived Stem Cells in a Scaffold System Attenuates Acute Liver Injury

doi: 10.5009/gnl18235

Figure Lengend Snippet: Stem cell homing. (A) Mouse and human β2-microglobulin mRNA expression was compared between the TAA-only group and the ADSCs scaffold group. (B) Immunofluorescence stain using human anti-mitochondria antibody was performed in mouse liver (×200) and implanted scaffold (×630) (immunofluorescence: blue - nuclei, green - human mitochondria). H&E staining (blue - nuclei, pink - cytosol, ×400) and periodic acid-Schiff (PAS) staining (blue - nuclei, purple - glycogen in cytoplasm, ×630) of implanted scaffold. TAA, thioacetamide; ADSCs, adipose tissue-derived stem cells; Foxa2, forkhead box protein.

Article Snippet: For liver function evaluation, periodic acid-Schiff staining (PAS Staining Kit, 1.01646.0001; Merck Millipore) was performed to analyze glycogen storage capacity.

Techniques: Expressing, Immunofluorescence, Staining, Derivative Assay

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