Journal: Journal of Clinical Investigation

Article Title: Loss of acinar cell IKKα triggers spontaneous pancreatitis in mice

doi: 10.1172/jci64498

Figure Lengend Snippet: Figure 2 Loss of pancreatic IKKα causes acinar cell death and compensa- tory proliferation. (A) Amylase and lipase activities in sera of 6- or 12-week-old IkkαF/F and IkkαΔpan mice. (B) Trypsin activity in pan- creata of 2- or 5-month-old IkkαF/F and IkkαΔpan mice. (C) TUNEL assay and apoptotic cell index. Original magnification, ×200. (D and E) H&E staining of IkkαΔpan pancreatic sections showing aci- nar cell apoptosis (D) and necrosis (E). Original magnification, ×400. (F) BrdU incorporation and label- ing index in pancreata of 3-month- old IkkαF/F and IkkαΔpan mice. Original magnification, ×400. (G) Ki67 staining and labeling index of pancreatic sections from 3-month- old mice. Original magnifica- tion, ×400. Results are shown as mean ± SEM. n = 4–6 mice per condition. **P < 0.01. Scale bars: 50 μm; 10 μm (insets).

Article Snippet: IB, IHC, and IF were conducted with antibodies to the following: IKKα (Imgenex), LC3B (Cell Signaling), p62 (Santa Cruz Biotechnology Inc. and Promgen), ERK (Cell Signaling), HA (Roche), Flag (Sigma-Aldrich), ATG16L2 (Abgent), Ub (Santa Cruz Biotechnology Inc.), LAMP1 and LAMP2 (Santa Cruz Biotechnology Inc.), Ki67 (Novocastra Laboratories), B220 (BD Biosciences — Pharmingen), CD3 (Abcam), F4/80 (AbD Serotec), PE-Gr1 (BD Biosciences — Pharmingen),insulin (Zymed), glucagon (Abcam), Sp1 (Santa Cruz Biotechnology Inc.), NF-κB/RelA (Santa Cruz Biotechnology Inc.), tubulin (Sigma-Aldrich), phospho-elF2α (Upstate), elF2α (Santa Cruz Biotechnology Inc.), phospho-ERK (Cell Signaling), JNK1/2 (Pharmingen), phospho-JNK (Cell Signaling), cathepsin B and cathepsin D (Santa Cruz Biotechnology Inc.), phospho-PERK (Cell Signaling), phospho-IKKα/β (Cell Signaling).

Techniques: Activity Assay, TUNEL Assay, Staining, BrdU Incorporation Assay, Labeling

Journal: Cells

Article Title: From Patient Liver Tissue to Organoids: Establishment of a Translational Platform Using Healthy, Steatotic, and Cirrhotic Tissue Sources

doi: 10.3390/cells15050432

Figure Lengend Snippet: Immunofluorescence imaging of patient-derived organoid culture. ( A ) ( a ) Overlay of all markers, ( b ) DAPI (blue, nuclear staining), ( c ) Ki-67 (cyan, proliferation marker), ( d ) Albumin (green, hepatocyte marker), ( e ) HNF4α (yellow, hepatocyte marker), ( f ) CK 19 (red, cholangiocyte marker), ( g ) LGR5 (pink, liver-specific stem-cell marker). ( B ) Healthy patients’ organoids ( a ) Overlay of all markers, ( b ) Collagen I (green, collagen marker), ( c ) SMA (red, hepatic stellate cells and myofibroblasts). ( C ) Cirrhotic patients organoids ( a ) Overlay of all markers, ( b ) Collagen I (green), ( c ) SMA (red). Scale bars: 200 µm.

Article Snippet: The human HNF4a qPCR primer pair (# HP200437 , NM_000457 , Origene), human MKI67 (# HP206104 , NM_002417.5 , Origene), human LGR5 ( NM_001277226.2 , [ ]), and human ALB ( NM_000477.7 , [ ]) were used.

Techniques: Immunofluorescence, Imaging, Derivative Assay, Staining, Marker

Journal: Cells

Article Title: From Patient Liver Tissue to Organoids: Establishment of a Translational Platform Using Healthy, Steatotic, and Cirrhotic Tissue Sources

doi: 10.3390/cells15050432

Figure Lengend Snippet: Analysis of antibody distribution in liver organoids derived from healthy, steatotic, and cirrhotic patients. The antibodies used were Ki-67, Albumin, HNF4α, CK19, Collagen I, and SMA. The bars in the figure represent the mean ± SD from three technical replicates (N = 3). An ordinary one-way ANOVA was conducted for statistical analysis (ns: non-significant, * p < 0.05).

Article Snippet: The human HNF4a qPCR primer pair (# HP200437 , NM_000457 , Origene), human MKI67 (# HP206104 , NM_002417.5 , Origene), human LGR5 ( NM_001277226.2 , [ ]), and human ALB ( NM_000477.7 , [ ]) were used.

Techniques: Derivative Assay

Journal: Cells

Article Title: From Patient Liver Tissue to Organoids: Establishment of a Translational Platform Using Healthy, Steatotic, and Cirrhotic Tissue Sources

doi: 10.3390/cells15050432

Figure Lengend Snippet: Relative expression of HNF4A , MKI67 , LGR5 , and ALB in human organoids as determined by qPCR. The bars represent the mean ± SD (N = 3). An ordinary one-way ANOVA was conducted for statistical analysis (* p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001).

Article Snippet: The human HNF4a qPCR primer pair (# HP200437 , NM_000457 , Origene), human MKI67 (# HP206104 , NM_002417.5 , Origene), human LGR5 ( NM_001277226.2 , [ ]), and human ALB ( NM_000477.7 , [ ]) were used.

Techniques: Expressing

Journal: Cell death & disease

Article Title: Cellular senescence promotes macrophage-to-myofibroblast transition in chronic ischemic renal disease.

doi: 10.1038/s41419-025-07666-1

Figure Lengend Snippet: Fig. 4 Senescent cells (SC) induce senescence in macrophages. A Schematic of the in vitro experimental protocol. B, C: Immunofluorescence staining of Ki67 and quantitative analysis of Ki67+ cells. The number of Ki67+ cells decreased after co-culture with senescent human renal proximal tubular epithelial cells (HRPTEpiC) (senescent cells, SC). Scale bar: 100 µm (B). The data are mean ± SD (n = 3/group). D–H: The effects of IFITM-3 and ITGB-3 on macrophage senescence. IFITM3 and ITGB3 were manipulated in SC HRPTEpiC and macrophages, respectively. Macrophages were subsequently collected for Western blot analysis of senescence markers p21Cip1/Waf1, p53, p-p53.S15, and γ-H2AX. Silencing ITGB3 or IFITM3 individually using siRNA reduced the expression of these senescence markers. However, this blunting effect was mitigated when IFITM3 was overexpressed in HRPTEpiC, indicating that IFITM3 plays a critical role in maintaining senescence signaling despite ITGB3 knockdown. Data are mean ± SD (n = 3/group). *P < 0.05 vs. Normal control (NC) and Non-SC groups; #P < 0.05 vs. SC group; and P < 0.05 vs. SC + IFITM3 over-expressing groups. I–K: Relative mRNA expression of the senescence markers p16INK-4a, p21Cip1/Waf1, and p53 increased in macrophages co-incubated with senescent cells. β-actin was used as loading control. Data are mean ± SD (n = 3/group). *P < 0.05 vs. Normal.

Article Snippet: For assessment of senescence by immunofluorescence, macrophages fixed with 4% formaldehyde were incubated with anti-CD68 (ab95564, Abcam), p16INK-4a (10883-1-AP, Protein tech), α-SMA (48938S, Cell Signaling), or Ki67 (AF7617, R&D Systems) and then a secondary antibody.

Techniques: In Vitro, Staining, Co-Culture Assay, Western Blot, Expressing, Knockdown, Control, Incubation

Journal: BMC Ophthalmology

Article Title: Protection effect of thymosin β4 on ethanol injury in corneal stromal keratocyte

doi: 10.1186/s12886-022-02255-8

Figure Lengend Snippet: A The results of ki67 on corneal tissue. Red showed the expression of Ki67, blue = DAPI. Scale bar: 500 μm. B Positive cell count of corneal stroma of four groups. * p < 0.05

Article Snippet: Sections of 4.5 μm were mounted onto cryostate (Leica CM 1850; Leica Microsystems GmbH, Wetzlar, Germany) and incubated with rabbit monoclonal antibody against ki67 (1:500, Catalogue # MAB7617, R&D Systems, MN, USA) at 4 °C overnight, followed by incubation with DyLight 594-conjugated goat anti-rabbit IgG (1:200, Thermo Fisher Scientific, Inc.) at room temperature for 1 h. Nuclei were stained with 4,6-diamidino-2-phenylindole (DAPI, 1:1000, Beijing Solarbio Science & Technology Co., Ltd., cat. no. S2110).

Techniques: Expressing, Cell Counting

Journal: Frontiers in cell and developmental biology

Article Title: Recreating the Bone Marrow Microenvironment to Model Leukemic Stem Cell Quiescence.

doi: 10.3389/fcell.2021.662868

Figure Lengend Snippet: FIGURE 2 | Bone marrow mesenchymal stromal cells (BMSC) and TGFβ-1 have an additive effect on slowing KG1a proliferation and lead to the accumulation of cells in the G0 cell cycle phase. KG1a cells were cultured with alginate-embedded BMSC with or without addition of 10 ng/ml TGFβ-1 for the times indicated. (A) Histogram representation of carboxyfluorescein succinimidyl ester (CFSE) retention in single culture at day 0 (red) and at day 4 or 8 (blue), culture with TGFβ-1 (orange), BMSC co-culture (green), or both (purple) on days 4 and 8. (B) Percentage of cells in the G0-, G1-, and S/G2-M phases of the cell cycle determined by Ki67 + PI staining. (C) Histogram representation of CFSE retention in extended single culture (blue) vs. BMSC + TGFβ-1 co-culture (purple). Red, CFSE fluorescence on day 0; US, unstained sample. *p < 0.05; ***p < 0.001 (Student’s t-test).

Article Snippet: Anti-CD150-PE, anti-GRPC5C Alexa Fluor-405, and anti-Ki67-Alexa Fluor 405 antibodies for flow cytometry were purchased from R&D Systems.

Techniques: Cell Culture, Co-Culture Assay, Staining

Journal: Frontiers in cell and developmental biology

Article Title: Recreating the Bone Marrow Microenvironment to Model Leukemic Stem Cell Quiescence.

doi: 10.3389/fcell.2021.662868

Figure Lengend Snippet: FIGURE 3 | Hypoxia enhances the quiescence-inducing effect of bone marrow mesenchymal stromal cells (BMSCs) and TGFβ-1. KG1a cells were labeled with 2.5 µM carboxyfluorescein succinimidyl ester (CFSE) and cultured alone (KG1a) or with alginate-encapsulated BMSC and TGFβ-1 (10 ng/ml) over a 12-day period in normoxia or hypoxia (1% pO2). (A) CFSE retention measured in the live cell fraction on days 1, 8, and 12. (B) Viability measured with Annexin V on days 1, 8, and 12. (C) The absolute number of live cells per milliliter as determined by flow cytometry using CountBright beads on days 1, 8, and 12. (D) Cell cycle distribution based on Ki67 expression and DNA content (PI) on days 8 and 12. *p < 0.05; **p < 0.01; ***p < 0.001 (Student’s t-test).

Article Snippet: Anti-CD150-PE, anti-GRPC5C Alexa Fluor-405, and anti-Ki67-Alexa Fluor 405 antibodies for flow cytometry were purchased from R&D Systems.

Techniques: Labeling, Cell Culture, Cytometry, Expressing