Journal: Discover oncology

Article Title: An integrative analysis combining bioinformatics, network pharmacology and experimental methods identified key genes of EGCG targets in Nasopharyngeal Carcinoma.

doi: 10.1007/s12672-025-02365-x

Figure Lengend Snippet: Fig. 7 To identify potential therapeutic targets for EGCG. A Expression of CYCS in the GEO cohort. B Visualized the distribution of the six- genes in the risk model within the previous volcano plot. C Expression of MYL12B in the GEO cohort

Article Snippet: Following this step, primary antibodies against CYCS (diluted 1:1000, #10,993–1-AP, Proteintech), and MYL12B (diluted 1:250, #10,324–1-AP, Proteintech) were applied, and the sections were left to incubate with the secondary antibody (kit-5020, MAIXIN BIOTECH, Fuzhou, China) for 70 min at room temperature.

Techniques: Biomarker Discovery, Expressing

Journal: Discover oncology

Article Title: An integrative analysis combining bioinformatics, network pharmacology and experimental methods identified key genes of EGCG targets in Nasopharyngeal Carcinoma.

doi: 10.1007/s12672-025-02365-x

Figure Lengend Snippet: Fig. 8 Immunohistochemistry of CYCS, and MYL12B. A, B CYCS protein levels in NPC tissue expression. C Diagnostic efficacy of CYCS in NPC based on protein level. D, E MYL12B protein levels in NPC tissue expression. F Diagnostic efficacy of MYL12B in NPC based on protein level

Article Snippet: Following this step, primary antibodies against CYCS (diluted 1:1000, #10,993–1-AP, Proteintech), and MYL12B (diluted 1:250, #10,324–1-AP, Proteintech) were applied, and the sections were left to incubate with the secondary antibody (kit-5020, MAIXIN BIOTECH, Fuzhou, China) for 70 min at room temperature.

Techniques: Immunohistochemistry, Expressing, Diagnostic Assay

Journal: Science Advances

Article Title: Inflammatory cytokines disrupt astrocyte exosomal HepaCAM-mediated protection against neuronal excitotoxicity in the SOD1G93A ALS model

doi: 10.1126/sciadv.adq3350

Figure Lengend Snippet: ( A ) Exosomes were isolated from untreated and ITC cytokine–treated primary NTg and SOD1G93A astrocyte cultures for LC-MS/MS proteomic analysis. Three-way Venn diagrams of proteins that were up-regulated ( B ) and down-regulated ( C ) in A-Exo. from astrocyte cultures treated with ITC cytokines and SOD1G93A expression. Fold change > or < 1.5 was used as the cutoff to identify changed proteins either by ITC treatment or SOD1G93A expression. The number of proteins that are overlapped in each comparison was highlighted in the diagram. Black arrows indicate either up- or down-regulated proteins in A-Exo from each comparison, presented as the numerator divided by the denominator. ( D ) Cellular location of proteins up-regulated with cytokine stimulation in NTg and SOD1G93A (co–up-regulated) and down-regulated with cytokine stimulation in NTg and SOD1G93A (co–down-regulated). ( E ) Heatmap of co–up-regulated and co–down-regulated plasma membrane proteins. Values are Log 2 (averaged iBAQ values). Representative immunoblot ( F ) and quantification ( G ) of HepaCAM protein in NTg, ITC-NTg, SOD1G93A, and ITC-SOD1G93A A-Exo. based on total loaded protein (gel staining intensity, 2 μg per lane); n = 5 to 6 independent samples per group. Representative HepaCAM immunoblot ( H ) and quantification ( I ) from lumbar spinal cords of SOD1G93A (onset, P90 to P100; mid-disease, P110 to P120) and age-matched NTg mice. N = 5 to 9 mice per group. β-Actin was used as the control for normalization. Error bars denote SEM, and P values are calculated using one-way ANOVA followed by a Tukey post hoc test in (G) and (I).

Article Snippet: This virus was packaged by the Boston Children’s Hospital Viral Core based on a human HepaCAM mGFP-tagged HepaCAM lentiviral vector (Origene Technologies, #RC223693L4V).

Techniques: Isolation, Liquid Chromatography with Mass Spectroscopy, Expressing, Comparison, Membrane, Western Blot, Staining, Control

Journal: Science Advances

Article Title: Inflammatory cytokines disrupt astrocyte exosomal HepaCAM-mediated protection against neuronal excitotoxicity in the SOD1G93A ALS model

doi: 10.1126/sciadv.adq3350

Figure Lengend Snippet: Cortical neuronal death rate ( A ) and neurite beading index ( B ) in primary cortical neurons treated with l -Glu (100 μM), NTg A-Exo. + l -Glu, HepaCAM KO A-Exo. + l -Glu, HepaCAM KO + HepaCAM overexpression (OE) A-Exo. + l -Glu. Control, 1× PBS; n = 24 to 31 fields (20×) per three biological replicates per group for neuronal death rate. n = 24 to 38 fields (20×) per three biological replicates per group for neurite beading analysis. ( C ) Representative images of βIII-TUBULIN and TUNEL staining in primary cortical neurons grown on PDL + BSA or PDL + HepaCAM ECD and treated with l -Glu. (100 μM). White arrows, neurites with beading; yellow arrows, preserved axons with minimal beading. (ii) A magnified view from the box in (i); (v) a magnified view from the box in (iv); circles indicate TUNEL + neurons in (iii). Scale bar, 30 μm [(i), (iii), (iv), and (vi)]; 20 μm [(ii) and (v)]. Cortical neuronal death rate ( D ) and neurite beading index ( E ) in primary cortical neurons grown on PDL + BSA or PDL + HepaCAM ECD and treated with l -Glu. (100 μM). n = 29 to 32 fields (20×) per three biological replicates per group for neuronal death rate. n = 18 to 26 fields (20×) per two biological replicates per group for neurite beading analysis. Data from different fields of the same biological replicate were averaged and presented in each panel. Error bars denote SEM, and P values in (A), (B), (D), and (E) were calculated using one-way ANOVA followed by a Tukey post hoc test.

Article Snippet: This virus was packaged by the Boston Children’s Hospital Viral Core based on a human HepaCAM mGFP-tagged HepaCAM lentiviral vector (Origene Technologies, #RC223693L4V).

Techniques: Over Expression, Control, TUNEL Assay, Staining

Journal: Science Advances

Article Title: Inflammatory cytokines disrupt astrocyte exosomal HepaCAM-mediated protection against neuronal excitotoxicity in the SOD1G93A ALS model

doi: 10.1126/sciadv.adq3350

Figure Lengend Snippet: ( A ) Schematic representation of the microfluidic chamber system and representative images of the soma and axonal compartments. (i) Differential interference contrast (DIC) image of neurons at the soma side; (ii) DIC image of axons at the axon side; (iii) tdT + -labeled neurons by adding AAV8-CAG-tdT onto the soma side of the chamber at 10 DIV; (iv) tdT + -labeled axons at the axon side of the chamber; white lines and gray arrows indicate the ending of the soma side (i) and the beginning of the axon side (ii) of the chamber. Scale bar, 100 μm. Representative images ( B ) of tdT + axon degeneration and quantification ( C ) of axonal tdT + intensity over time following addition of l -Glu. into either soma or axon sides. White arrows, degenerating axons indicated by beading following soma side of l -Glu. treatment; yellow arrows, healthy axons even after l -Glu. treatment at the axon side; scale bar, 50 μm; n = 19 to 30 fields (10×) per three chambers per two biological replicates per group. Diagram of the experiment and representative tdT + axon images ( D ) and tdT + intensity quantification ( E ) at the axon side of the microfluidic chambers. l -Glu was added onto the soma side and all A-Exo. (NTg and HepaCAM A-Exo.) were added onto the axon side. White lines and gray arrows indicate the beginning of the axon side of the chamber. HepaCAM ECD was coated on microfluidic chambers. Time-lapse tdT + images were taken at 0-, 6-, and 22-hours after l -Glu. treatment, and the tdT fluorescent intensity was quantified. n = 19 to 39 fields (10×) per four chambers per two biological replicates per group; white arrows, degenerating axons; yellow arrows, healthy axons; scale bar, 50 μm. Error bars denote SEM. P values were calculated using two-way ANOVA followed by a Tukey post hoc test.

Article Snippet: This virus was packaged by the Boston Children’s Hospital Viral Core based on a human HepaCAM mGFP-tagged HepaCAM lentiviral vector (Origene Technologies, #RC223693L4V).

Techniques: Labeling

Journal: bioRxiv

Article Title: Growth Factor-Based Manufacturing of Human Pluripotent Stem Cell-Derived Cardiomyocytes Using the Vertical Wheel Bioreactor System

doi: 10.1101/2025.09.07.674777

Figure Lengend Snippet: A) Schematic of the protocols used to produce either ventricular CMs or atrial CMs. The ventricular differentiation protocol (referred to as 10B:6A) involves mesoderm induction with 10 ng/mL BMP4 and 6 ng/mL activin A from days 1-3, while the atrial protocol (referred to as 4B:2A) involves mesoderm induction using 4 ng/mL BMP4 and 2ng/mL activin A as well as addition of 0.5 µM retinoic acid (RA) from day 3-5. Cells were assessed for mesoderm induction efficiency using PDGFRα and CD56 on day 3. On day 4, ALDH activity and CD235a expression were used to identify second heart field (SHF) and first heart field (FHF) populations. B) Representative mesoderm profile at day 3 and C) day 4 of the differentiation protocol. D) Representative images of aggregate morphology on days 6 and 12 following differentiation with either the ventricular or atrial protocol. E) Representative flow cytometry plots for pan-cardiac marker cTnT and ventricular marker MLC2v in day 20 cultures generated with each differentiation protocol. Quantification of the percentage of cells in each condition expressing F) cTnT + , G) MLC2v, and H) MLC2a. I) Expression of ventricular or atrial genes at day 20 of differentiation. *p<0.05 by two-tailed Student’s t-test.

Article Snippet: To assess hPSC-CM purity, hPSC-CMs were fixed with 4% (w/v) paraformaldehyde (PFA) for 10 minutes, permeabilized with ice-cold 90% methanol for 20 minutes, and stained with primary antibodies against cardiac troponin T (cTnT, Miltenyi Biotec), myosin light chain-2v (MLC2v, Miltenyi Biotec), and myosin light chain-2a (MLC2a, Miltenyi Biotec) for 30 minutes at room temperature.

Techniques: Activity Assay, Expressing, Flow Cytometry, Marker, Generated, Two Tailed Test

Journal: bioRxiv

Article Title: Cryopreservation of human pluripotent stem cell-derived cardiomyocytes is not detrimental to their molecular and functional properties

doi: 10.1101/700849

Figure Lengend Snippet: A) Percentage of viable cells directly after dissociation ( fresh ) or upon thawing ( cryopreserved ); n=14 from 7 independent differentiations. B) Immunofluorescence image of cryopreserved hiPSC-CMs 6 days after thawing following staining with the sarcomere markers, α-actinin (red) and myosin heavy chain (MHC, green). Nuclei were stained with DAPI (blue). C) Recovery of fresh (blue) and cryopreserved (red) hiPSC-CMs at day 1, 4 and 7/8 post-replating. * indicates statistical significance (day 1 p=0.011, day 7/8 p=0.007, unpaired t -test); n=8 from 4 independent differentiations D) Representative flow cytometric analysis of d21+7 hiPSC-CMs for expression of cTnT, MLC2a and MLC2v. Left column is non-frozen CMs, while the remaining columns are cryopreserved hiPSC-CMs plated at either the same density (0.9×10 5 /cm 2 ; centre column ) or two-fold higher density (1.8×10 5 /cm 2 ; right column ) as the non-frozen CMs. Top row depicts histogram plots of cTnT expression, while remaining rows depict bivariate density plots of MLC2a/cTnT ( middle row ) and MLC2v/cTnT ( bottom row ). Numbers inside the plots are percentage of cells within the gated region. Dotted lines represent a control cTnT - population. E) Bar graph showing difference (D) in cTnT expression compared to the corresponding fresh hiPSC-CMs when the cryopreserved hiPSC-CMs were seeded at low (0.9×10 5 /cm 2 ) or high (1.8-2.1×10 5 /cm 2 ) density. * indicates statistical significance (p=0.016, unpaired t -test) (n=4) F) Bar graph showing percentage of cryopreserved cTnT + hiPSC-CMs expressing MLC2a or MLC2v when seeded at either low (0.9×10 5 /cm 2 ) or high (1.8×10 5 /cm 2 ) density (n=4) G) Bar graph showing percentage of freshly replated and cryopreserved cells expressing cTnT, as well as proportion of cTnT + hiPSC-CMs expressing MLC2a or MLC2v after taking into account differences in replating efficiencies for both LUMC20 ( left ) and LUMC99 ( right ) (n=7 and n=4 respectively).

Article Snippet: Cells were co-labelled with cTnT (Vioblue-conjugated), MLC2a (APC- or FITC-conjugated), and MLC2v (PE-conjugated) antibodies (all Miltenyi).

Techniques: Immunofluorescence, Staining, Expressing, Control