Journal: iScience

Article Title: Mutation T9I in Envelope confers autophagy resistance to SARS-CoV-2 Omicron

doi: 10.1016/j.isci.2025.112974

Figure Lengend Snippet: E T9I has increased affinity to autophagosome-associated proteins (A) Principal component analysis of the differential interactome data , the individual replicates are separated (black: GFP controls, Green: E T9 pulldown, Purple: E T9I pulldown) (B) Volcano plot of the differential interactome analysis showing enriched proteins in E T9I pulldown versus the p value (-log P). Five highly significantly enriched proteins are highlighted in red and via labels. (C–G) Quantification of proximity ligation assays between transiently expressed SARS-CoV-2 E variants 30 h post transfection in HeLa cells and endogenous SNX12, STX12, TMEM87B, ABCG2 and TAB1, as indicated. Representative images depicted. PLA signal, red. Scale Bar, 10μm. DAPI, nuclei (blue). Lines represent the mean of N = 18–59 (individual cells) ±SEM. (H) Quantification of autophagosome levels by flow cytometry in HEK293T autophagy reporter cells (HEK293T-GL) transiently expressing StrepII-tagged SARS-CoV-2 E variants (48 h post transfection) and depleted of indicated proteins by siRNA. Bars represent the mean of N = 3 (biological replicates) ±SEM. Student’s t test with Welch’s correction. ∗, p < 0.05; ∗∗, p < 0.01; ∗∗∗, p < 0.001.

Article Snippet: Monoclonal rabbit anti-ABCG2 Antibody (2K8X1) (WB 1:500) , Novus Biologicals , Cat#NBP3-15559.

Techniques: Ligation, Transfection, Flow Cytometry, Expressing

Journal: BioMed Research International

Article Title: Effects of Baicalin on Alopecia and the Associated Mechanism

doi: 10.1155/2022/3139123

Figure Lengend Snippet: Effects of baicalin or LY294002 in vitro . (a) HHDPC viability following baicalin treatment. (b) Effect of baicalin or LY294002 on HHDPC viability. (c) Representative images of the different HHDPC groups (scale bar, 100 μ m). (d) Western blot for the detection of the protein levels of AKT, p-AKT, IGF1, and ALP and (e) quantified relative expression values/ratios. ∗ P < 0.05 and ∗∗ P < 0.01 vs. the control group; # P < 0.05, ## P < 0.01, and ### P < 0.001 vs. the baicalin group. HHDPC: human hair dermal papilla cell; IGF1: insulin-like growth factor-1; ALP: alkaline phosphatase; p-AKT: phosphorylated AKT; OD: optical density.

Article Snippet: Primary antibodies AKT (1 : 1000, ab283852, Abcam, UK), phosphorylated p-AKT (1 : 500, ab8805, Abcam, UK), insulin-like growth factor-1 (IGF1, 1 : 1000, ab182408, Abcam, UK), alkaline phosphatase (ALP, 1 : 1000, ab229126, Abcam, UK), and β -actin (1 : 7000, ab8227, Abcam, UK) were used for incubation overnight at 4°C.

Techniques: In Vitro, Western Blot, Expressing, Control

Journal: Stem cell research

Article Title: IL6 and the human limbal stem cell niche: a mediator of epithelial-stromal interaction.

doi: 10.1016/j.scr.2010.07.002

Figure Lengend Snippet: Figure 4 (A and B) p63α (TRITC) and ABCG2 (TRITC) expression by HLE under 3.1SF conditions. Cytokeratin 3 is expressed by enlarging epithelial cells (B, FITC). (C) Semiquantitative RT-PCR data confirm the immunostaining observations. CFE assay of HLE cells derived by (D) GS and (E) 3.1SF cultures. Percentage CFE is significantly increased in 3.1SF cultures compared to GS (P=0.0286, m=6).

Article Snippet: Primary antibodies were supplied as follows: mouse monoclonal antibody for cytokeratin 15, Santa Cruz, mouse monoclonal antibody for ABCG2, mouse monoclonal antibody for cytokeratin 3 and mouse monoclonal antibody for Ki67, Millipore; rabbit polyclonal antibody for p63α, Cell Signalling, and rabbit polyclonal antibody for IL6, Abcam.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Immunostaining, Derivative Assay

Journal: Biomaterials Research

Article Title: Repairing Effect and Mechanism of the 4-Dimensionally Printed Limbal Stem Cell Strategy on Corneal Alkali Burns in Large Animals

doi: 10.34133/bmr.0262

Figure Lengend Snippet: Identification of the stemness of porcine limbal stem cells (PLSCs) and evaluation of the biocompatibility of 4D-CTH. (A) Representative images of primary culture of PLSCs at different time points. (B) Dual fluorescence staining of cytokeratin 14 (CK14) and Delta Np63 alpha (ΔNp63α) to identify primary PLSCs. (C) Dual fluorescence staining of B lymphoma Mo-MLV insertion region 1 homolog (BMI1) and ATP-binding cassette sub-family G member 2 (ABCG2) to identify primary PLSCs. (D) Migration ability of 4D-CTH-encapsulated limbal stem cells (LSCs) in the flat scratch assay. (E) Representative images of live and dead cell staining of 4D-CTH- and CTH-encapsulated LSCs. (F) Quantitative analysis of healing rate in the flat scratch assay. (G) Quantitative analysis of cell viability after 4D-CTH- and CTH-encapsulated LSCs. Scale bar, 50 μm; white scale bar, 200 μm. ns, not significant. D0, D3, D5, and D7, days 0, 3, 5, and 7; DAPI, 4′,6-diamidino-2-phenylindole.

Article Snippet: After sealing, the plates were fixed with 4% paraformaldehyde fixative (Sigma-Aldrich, USA) at room temperature for 20 min, followed by permeabilization with 0.2% Triton X-100 (Sigma-Aldrich, USA) for 5 min. After blocking for 30 min, the following specific primary antibodies were added and incubated overnight at 4 °C: BMI1 antibody (Cell Signaling Technology, USA), ΔNp63α antibody (Cell Signaling Technology, USA), CK14 antibody (Cell Signaling Technology, USA), and ABCG2 antibody (Proteintech, China).

Techniques: Fluorescence, Staining, Binding Assay, Migration, Wound Healing Assay

Journal: PLoS ONE

Article Title: Characterization of Cell Subpopulations Expressing Progenitor Cell Markers in Porcine Cardiac Valves

doi: 10.1371/journal.pone.0069667

Figure Lengend Snippet: Cell Surface Protein Markers.

Article Snippet: The cells were subsequently stained with selected primary antibodies raised against ABCG2 (R&D Systems, Cat # FAB995P), CD31 (AbD Serotec, Cat# MCA1746APC), OB–CDH (clone 15F7 [ ], a generous gift from Dr. Micheal Brenner’s group at the Brigham and Women’s Hospital) and SSEA4 (R&D Systems, Cat# FAB1435P) for 1 hour at 4°C.

Techniques:

Journal: PLoS ONE

Article Title: Characterization of Cell Subpopulations Expressing Progenitor Cell Markers in Porcine Cardiac Valves

doi: 10.1371/journal.pone.0069667

Figure Lengend Snippet: Freshly isolated aortic pVICs were stained with antibodies for CD31, OB–CDH, ABCG2, NG2, SSEA-4, and the corresponding control antibodies. Staining was quantified by flow cytometry. In the figure, the y-axis is fluorescence intensity and the x-axis is forward scattering. Percentage in the rectangular gates represents the fraction of positively stained cells. After subtracting the background, about 7.70% of these aortic pVICs stained positive for CD31, 4.71% stained positive for OB–CDH, 5.60% stained positive for ABCG2, 5.56% stained positive for NG2, and 6.59% stained positive for SSEA-4.

Article Snippet: The cells were subsequently stained with selected primary antibodies raised against ABCG2 (R&D Systems, Cat # FAB995P), CD31 (AbD Serotec, Cat# MCA1746APC), OB–CDH (clone 15F7 [ ], a generous gift from Dr. Micheal Brenner’s group at the Brigham and Women’s Hospital) and SSEA4 (R&D Systems, Cat# FAB1435P) for 1 hour at 4°C.

Techniques: Isolation, Staining, Control, Flow Cytometry, Fluorescence

Journal: PLoS ONE

Article Title: Characterization of Cell Subpopulations Expressing Progenitor Cell Markers in Porcine Cardiac Valves

doi: 10.1371/journal.pone.0069667

Figure Lengend Snippet: Unique cell subpopulations identified based on cell surface markers are present in both aortic and pulmonary valves.

Article Snippet: The cells were subsequently stained with selected primary antibodies raised against ABCG2 (R&D Systems, Cat # FAB995P), CD31 (AbD Serotec, Cat# MCA1746APC), OB–CDH (clone 15F7 [ ], a generous gift from Dr. Micheal Brenner’s group at the Brigham and Women’s Hospital) and SSEA4 (R&D Systems, Cat# FAB1435P) for 1 hour at 4°C.

Techniques: Marker

Journal: PLoS ONE

Article Title: Characterization of Cell Subpopulations Expressing Progenitor Cell Markers in Porcine Cardiac Valves

doi: 10.1371/journal.pone.0069667

Figure Lengend Snippet: To examine co-expression of these different markers, freshly isolated pVICs were co-stained with pairwise combinations of the markers OB–CDH, NG2 and SSEA-4, or with ABCG2 and NG2. Both the x-axes and the y-axes are fluorescence intensity of the antibody staining. (A) OB–CDH was not co-expressed with SSEA-4 in the same cell population. (B) OB–CDH was not co-expressed with NG2 in the same cell population. (C) SSEA-4 was not co-expressed with NG2 in the same cell population. These data support that these markers (OB–CDH, NG2 and SSEA-4) label distinct subpopulations relative to each other in pVICs. (D) However, NG2 and ABCG2 were co-expression by the same cell subpopulation in aortic valves.

Article Snippet: The cells were subsequently stained with selected primary antibodies raised against ABCG2 (R&D Systems, Cat # FAB995P), CD31 (AbD Serotec, Cat# MCA1746APC), OB–CDH (clone 15F7 [ ], a generous gift from Dr. Micheal Brenner’s group at the Brigham and Women’s Hospital) and SSEA4 (R&D Systems, Cat# FAB1435P) for 1 hour at 4°C.

Techniques: Expressing, Isolation, Staining, Fluorescence

Journal: PLoS ONE

Article Title: Characterization of Cell Subpopulations Expressing Progenitor Cell Markers in Porcine Cardiac Valves

doi: 10.1371/journal.pone.0069667

Figure Lengend Snippet: To examine the origin of these cell subpopulations, aortic pVICs were double stained with CD31 and one of the following markers: SSEA-4, OB–CDH or ABCG2. CD31 is expressed by porcine valvular endothelial cells but not by pVICs. From (A), SSEA4 and CD31 labeled distinct cell subpopulations. However, OB–CDH + or ABCG2 + cells showed heterogeneous CD31 staining. About 52% of OB–CDH + cells were CD31 - and ~61% of ABCG2 + cells were CD31 - , indicating that a majority of these cells are from pVICs rather than endothelium.

Article Snippet: The cells were subsequently stained with selected primary antibodies raised against ABCG2 (R&D Systems, Cat # FAB995P), CD31 (AbD Serotec, Cat# MCA1746APC), OB–CDH (clone 15F7 [ ], a generous gift from Dr. Micheal Brenner’s group at the Brigham and Women’s Hospital) and SSEA4 (R&D Systems, Cat# FAB1435P) for 1 hour at 4°C.

Techniques: Staining, Labeling

Journal: PLoS ONE

Article Title: Characterization of Cell Subpopulations Expressing Progenitor Cell Markers in Porcine Cardiac Valves

doi: 10.1371/journal.pone.0069667

Figure Lengend Snippet: Sorted ABCG2 + and ABCG2 - valvular cells were grown to confluence and re-plated in osteogenic media for 8 days. (A) Brightfield images of cells cultured in control conditions or osteogenic conditions were taken at day 8. ABCG2 + progeny secreted more dark mineralization spots than ABCG2 - progeny on plastic plates. Scale bar: 100 µm. (B) Calcium deposition by the cells was quantified as described in the Materials and Methods. Data was represented as mean + standard error. Compared with ABCG2 - progeny, ABCG2 + progeny produced higher amounts of calcium composites at day 8 (* indicates p<0.05 in between the groups).

Article Snippet: The cells were subsequently stained with selected primary antibodies raised against ABCG2 (R&D Systems, Cat # FAB995P), CD31 (AbD Serotec, Cat# MCA1746APC), OB–CDH (clone 15F7 [ ], a generous gift from Dr. Micheal Brenner’s group at the Brigham and Women’s Hospital) and SSEA4 (R&D Systems, Cat# FAB1435P) for 1 hour at 4°C.

Techniques: Cell Culture, Control, Produced

Journal: PLoS ONE

Article Title: Characterization of Cell Subpopulations Expressing Progenitor Cell Markers in Porcine Cardiac Valves

doi: 10.1371/journal.pone.0069667

Figure Lengend Snippet: Based on previous literature and our flow cytometry data, porcine aortic valves are comprised of layers of endothelial cells and VICs. pVICs are further divided into multiple different subpopulations, including ABCG2 + and NG2 + pVICs, OB–CDH + pVICs and SSEA-4 + pVICs. The functions of these different subpopulations in the diseased valves are speculated based on literature and our experimental data, and these serve as hypotheses to be tested in future.

Article Snippet: The cells were subsequently stained with selected primary antibodies raised against ABCG2 (R&D Systems, Cat # FAB995P), CD31 (AbD Serotec, Cat# MCA1746APC), OB–CDH (clone 15F7 [ ], a generous gift from Dr. Micheal Brenner’s group at the Brigham and Women’s Hospital) and SSEA4 (R&D Systems, Cat# FAB1435P) for 1 hour at 4°C.

Techniques: Flow Cytometry

Journal: Pharmaceutics

Article Title: Targeted Self-Emulsifying Drug Delivery Systems to Restore Docetaxel Sensitivity in Resistant Tumors.

doi: 10.3390/pharmaceutics14020292

Figure Lengend Snippet: Figure 3. Enox-coated SEDDS inhibit MRP1 and BCRP activity. A459 cells were incubated for (a) 24 h or (c) 72 h with fresh medium (ctrl), 80 µM free docetaxel (DTX), 0.25% v/v blank SEDDS (S), Enox-coated SEDDS (S/Enox-Pa), SEDDs containing docetaxel (80 µM final concentration; S/DTX), and Enox-coated SEDDS containing docetaxel (80 µM final concentration; S/Enox-Pa/DTX). (a) The rate of ATP hydrolysis by immunopurified Pgp, MRP1, or BCRP extracted from cells treated as reported above was measured by spectrophotometric analysis in triplicates. Data are presented as means + SD (n = 3). * p < 0.02: vs. ctrl. (b) A549 cells were transduced with a non-targeting (scrambled) CRISPR-Cas vector or with a CRISPR-Cas vector to knock-out (KO) MRP1 or BCRP. The indicated proteins were measured by immunoblotting. Tubulin was used as control of equal protein loading. The image is representative of one out of three experiments. (c,d) The viability of scrambled, KO MRP1, and KO BCRP A549 cells was measured by a chemiluminescence-based assay in quadruplicates. Data are presented as means + SD (n = 3). * p < 0.001: vs. ctrl; ◦p < 0.001: S/Enox-Pa/DTX vs. DTX; # p < 0.01: S/Enox-Pa/DTX vs. S/DTX; § p < 0.001: KO vs. scrambled cells.

Article Snippet: The non-targeting siRNA sequence (Trilencer-27 Universal scrambled negative control siRNA duplex, #R30004), the FGFR1-targeting siRNAs pool of 3 unique 27mer siRNA duplexes (#SR320159), the CRISPR pCas vectors targeting MRP1 (#KN418182), BCRP (#KN405640), or the non-targeting vector (#GE100003) were purchased from Origene (Rockville, MD, USA).

Techniques: Activity Assay, Incubation, Concentration Assay, Transduction, CRISPR, Plasmid Preparation, Knock-Out, Western Blot, Control, Chemiluminescence Immunoassay

Journal: Journal of cell communication and signaling

Article Title: Quinacrine and Curcumin in combination decreased the breast cancer angiogenesis by modulating ABCG2 via VEGF A.

doi: 10.1007/s12079-022-00692-0

Figure Lengend Snippet: Fig. 6 Schematic representation of ABCG2 induced angiogenesis. VEGF A from ABCG2 enriched SP cells interacts with VEGF R and activates tumor induced angiogenesis via PI3K, Akt and eNOS

Article Snippet: ABCG2 siRNA (#sc-41,151) was purchased from Santa Cruz Biotechnology Inc., CA, USA.

Techniques:

Journal: Cancers

Article Title: Identification of EZH2 as Cancer Stem Cell Marker in Clear Cell Renal Cell Carcinoma and the Anti-Tumor Effect of Epigallocatechin-3-Gallate (EGCG).

doi: 10.3390/cancers14174200

Figure Lengend Snippet: Figure 3. The sphere formation ability of the RCC cell lines and the expression of potential CSC mark- ers on the mRNA and protein levels. (A) Sphere formation: SKRC-17 and RCC-53 formed spheres around day 7, RCC-26 did not. Photos were taken by a microscope digital camera at the magnification of 100× (Bresser GmbH DE-46414 Rhede Germany). (B) mRNA expression of 19 potential CSC markers in SKRC-17, RCC-53, and their corresponding CSCs analyzed by RT-qPCR analysis (p < 0.05 indicates statistical significance), the expression level observed in spheres was normalized to the corresponding adherent line. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. (C) Representative images of IHC staining for the EZH2, ABCG2, ALDH1A3, and SALL4 expression in the KIRC and normal tissue, photos derived from the Human Protein Atlas database. (D) Measurement of the protein expression in the adherent and sphere cell lines by flow cytometry (n = 3, adherent cell value was set as 1).

Article Snippet: Spheres and adherent tumor cell lines were harvested and stained with the following mouse monoclonal antibodies: EZH2 (clone 11/EZH2 Alexa Fluor® 647, BD Biosciences, Heidelberg, Germany), ALDH1A3 (clone OTI4E8, ORIGENE, Rockville, MD, USA), SALL4 (clone 6E3, Abcam, Cambridge, UK), and ABCG2 (clone 5D3/CD338, APC, BD Biosciences), respectively.

Techniques: Expressing, Microscopy, Quantitative RT-PCR, Immunohistochemistry, Derivative Assay, Cytometry