Journal: Materials Today Bio

Article Title: Injectable chitosan-based hydrogel via in situ gelation modulates the inflammatory microenvironment and facilitates minimally invasive repair of peripheral nerve injury

doi: 10.1016/j.mtbio.2026.102814

Figure Lengend Snippet: Pro-angiogenic and pro-migratory effects of hydrogels. Immunofluorescence staining shows blood vessel formation of HUVECs in LPS-macrophage condition medium (A). Scratch assays and Transwell migration assays of HUVECs (B) and L929 cells (C). Quantification of junctions (D), branches (E), wound closure percentage (F–G) and migrated cells (H–I). One-way ANOVA with Tukey's post hoc test and n = 3 for D-I; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, n.s. ANOVA, analysis of variance; CS, chitosan; IBU, ibuprofen; GP, genipin; MA, methacrylic anhydride; LPS, lipopolysaccharides; HUVEC, human umbilical vein endothelial cell; n.s., not significant.

Article Snippet: Mouse fibroblasts (L929, ATCC), human umbilical vein endothelial cells (HUVECs, ATCC), and mouse macrophages (RAW 264.7, ATCC) were provided by the Cell Bank of the Chinese Academy of Sciences.

Techniques: Immunofluorescence, Staining, Migration

Journal: Regenerative Therapy

Article Title: Airway basal stem cell derived extracellular vesicles promote lung repair in chronic obstructive pulmonary disease

doi: 10.1016/j.reth.2026.101068

Figure Lengend Snippet: Characterization of BC-EVs. (a) Representative transmission electron microscopy image of BC-EVs. Scale bar, 100 nm. (b) Size distribution of BC-EVs determined by Nano-Flow Cytometry. (c) Western blotting analysis showing the expression of EV markers TSG101 and CD63, and the negative marker Calnexin. (d) Representative images of HUVECs tube formation after treatment with DMEM, BC-derived EVs, or MSC-derived EVs. Scale bar, 100 μm. (e) Quantification of the number of tubes formed in each group (n = 5 technical replicates). (f) Cell viability of BCs cultured under serum-depleted conditions and treated with DMEM, or increasing concentrations of BC-EVs or MSC-EVs (n = 3 technical replicates). (e) and (f) All data are presented as mean ± SEM. Statistical analysis was performed using a one-way ANOVA with Tukey's multiple comparisons test. ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001; ns, not significant.

Article Snippet: Human umbilical vein endothelial cells (HUVECs) were obtained from the American Type Culture Collection (ATCC) and cultured in DMEM supplemented with 10 % fetal bovine serum.

Techniques: Transmission Assay, Electron Microscopy, Flow Cytometry, Western Blot, Expressing, Marker, Derivative Assay, Cell Culture

Journal: NAR Cancer

Article Title: Ribosome biogenesis is increased in hepatocellular carcinoma and represents a potential therapeutic target

doi: 10.1093/narcan/zcaf058

Figure Lengend Snippet: Nucleolar factors associated with FC/DFC are upregulated at both mRNA and protein level in HCC. Relative mRNA levels of ( A ) TCOF1 (Treacle), ( B ) UBF, ( C ) FBL (Fibrillarin), ( D ) NCL (nucleolin), and ( E ) NPM1 (nucleophosmin) measured by qPCR in HCC cells (PLC/PRF/5 and SNU-449) and control cells (THLE-3). GAPDH was used as the housekeeping gene, and fold change was determined relative to mRNA levels in THLE-3 cells. Individual points in the graph represent biological replicates ( n = 3). The graph depicts the mean ± SD, with statistical significance assessed by one-way ANOVA and indicated with numerical P -values. Protein levels of ( F ) Treacle, ( G ) UBF, ( H ) Fibrillarin, ( I ) nucleolin, and ( J ) nucleophosmin measured by WB in HCC cells (PLC/PRF/5 and SNU-449) and control cells (THLE-3). α-tubulin was used as a loading control (LC). The molecular weight in kDa is listed on the left side of the blots (left panel), and protein signals were determined in each cell line relative to the loading control (right panel). Individual points in the graphs represent biological replicates ( n = 3). The graph depicts the mean ± SD, with statistical significance assessed by one-way ANOVA and indicated by numerical P -values.

Article Snippet: Human osteosarcoma U2OS cells (#HTB-96), HCC cell lines PLC/PRF/5 (#CRL-8024), SNU-449 (#CRL-2234), and normal liver epithelial cells THLE-3 (#CRL-3583) were purchased from the American Type Culture Collection.

Techniques: Control, Molecular Weight

Journal: NAR Cancer

Article Title: Ribosome biogenesis is increased in hepatocellular carcinoma and represents a potential therapeutic target

doi: 10.1093/narcan/zcaf058

Figure Lengend Snippet: Nucleolar expression of FC/DFC-associated factors is upregulated in HCC. Representative images of ( A ) Treacle (green), ( B ) UBF (green), ( C ) Fibrillarin (green), and ( D ) nucleolin (green) and nucleophosmin (magenta), and Pol II (yellow) in THLE-3, PLC/PRF/5, and SNU-449 cells. ( E ) Total nucleolar Treacle intensity per nucleus. Graphs depict one representative replicate ( n = 3) with data points representing individual cells and the median indicated by the red line. A Kruskal–Wallis test was applied to assess differences between groups for all three biological replicates combined, and statistical significance is indicated by numerical P -values. ( F ) Total nucleolar UBF intensity per nucleus otherwise as in panel (E). ( G ) Total nucleolar Fibrillarin intensity per nucleus, otherwise as in panel (E). ( H ) Total nucleolar nucleolin intensity per nucleus otherwise as in panel (E). (I) Total nucleolar nucleophosmin intensity per nucleus otherwise as in panel (E).

Article Snippet: Human osteosarcoma U2OS cells (#HTB-96), HCC cell lines PLC/PRF/5 (#CRL-8024), SNU-449 (#CRL-2234), and normal liver epithelial cells THLE-3 (#CRL-3583) were purchased from the American Type Culture Collection.

Techniques: Expressing

Journal: NAR Cancer

Article Title: Ribosome biogenesis is increased in hepatocellular carcinoma and represents a potential therapeutic target

doi: 10.1093/narcan/zcaf058

Figure Lengend Snippet: rDNA transcription is increased in HCC. ( A ) Representative images of EU incorporation (green). THLE-3, PLC/PRF/5, and SNU-449 cells were stained with DAPI (blue) and an antibody against Pol II (yellow). ( B ) Total nucleolar EU intensity per nucleus as a readout for rDNA transcription in THLE-3, PLC/PRF/5, and SNU-449 cells. The graph depicts one representative biological replicate ( n = 3), with data points representing individual cells, and the median indicated by the red line. A Kruskal–Wallis test was applied to assess differences between groups for the replicates combined, and statistical significance is indicated by numerical P -values. ( C ) Nucleolar size measured through an inverse intensity-based mask of Pol II in THLE-3, PLC/PRF/5, and SNU-449 cells. Individual points in the graph represent biological replicates ( n = 3). The graph depicts the mean ± SD, and statistical significance assessed by one-way ANOVA is indicated with numerical P -values. ( D ) Number of nucleoli per nucleus in THLE-3, PLC/PRF/5, and SNU-449 cells measured through an inverse intensity-based mask of Pol II. Individual points in the graph represent biological replicates ( n = 3). The graph depicts the mean ± SD, and statistical significance assessed by one-way ANOVA is indicated with numerical P -values. ( E ) Total nucleolar area otherwise as in panel (C).

Article Snippet: Human osteosarcoma U2OS cells (#HTB-96), HCC cell lines PLC/PRF/5 (#CRL-8024), SNU-449 (#CRL-2234), and normal liver epithelial cells THLE-3 (#CRL-3583) were purchased from the American Type Culture Collection.

Techniques: Staining

Journal: NAR Cancer

Article Title: Ribosome biogenesis is increased in hepatocellular carcinoma and represents a potential therapeutic target

doi: 10.1093/narcan/zcaf058

Figure Lengend Snippet: Anti-cancer drugs targeting the nucleolus inhibit cell viability in HCC. Drug response curves with corresponding IC 50 values and 95% CIs. Drugs were applied for 72 h, after which cell viability was assessed in THLE-3, PLC/PRF/5, and SNU-449 cells. ( A ) Drug response curve for CX-5461 with obtained IC 50 values and 95% CI (left panel). The graph shows the obtained IC 50 values represented as the mean ± SD (right panel). Individual points in the graph represent biological replicates ( n = 3). Statistical significance was assessed by one-way ANOVA and are indicated with numerical P -values. ( B ) Drug response curve for BMH-21 otherwise as in panel (A). ( C ) Drug response curve for Oxaliplatin, otherwise as in panel (A). ( D ) Drug response curve for JP-1302 otherwise as in panel (A). ( E ) Drug response curve for Sorafenib otherwise as in panel (A).

Article Snippet: Human osteosarcoma U2OS cells (#HTB-96), HCC cell lines PLC/PRF/5 (#CRL-8024), SNU-449 (#CRL-2234), and normal liver epithelial cells THLE-3 (#CRL-3583) were purchased from the American Type Culture Collection.

Techniques:

Journal: NAR Cancer

Article Title: Ribosome biogenesis is increased in hepatocellular carcinoma and represents a potential therapeutic target

doi: 10.1093/narcan/zcaf058

Figure Lengend Snippet: Nucleolar-targeting compounds inhibit nucleolar activity and induce selective DNA damage. ( A ) Representative images of EU incorporation (green) in THLE-3, PLC/PRF/5, and SNU-449 cells 24 h after treatment with four anti-cancer drugs targeting the nucleolus: CX-5461, BMH-21, Oxaliplatin, and JP-1302, or targeted therapy Sorafenib. Cells were treated with DMSO as control and were stained with DAPI (blue) and Pol II (yellow). ( B ) Mean nucleolar EU intensity per nucleus as a readout for rDNA transcription in THLE-3, PLC/PRF/5, and SNU-449 cells 24 h after treatment with four anti-cancer drugs targeting the nucleolus: CX-5461, BMH-21, Oxaliplatin, and JP-1302, or targeted therapy Sorafenib. Cells were treated with DMSO as control. The graph depicts one representative biological replicate ( n = 3), with data points representing individual cells, and the median indicated by the red line. A Kruskal–Wallis test was applied to assess differences between groups for the replicates combined, and statistical significance is indicated by numerical P -values. ( C ) Representative images of nuclear 53BP1 and γH2AX foci accumulation in THLE-3, PLC/PRF/5, and SNU-449 cells 24 h after treatment with four anti-cancer drugs targeting the nucleolus: CX-5461, BMH-21, Oxaliplatin, and JP-1302, or targeted therapy Sorafenib. Cells were treated with DMSO as control, and stained with DAPI (blue), 53BP1 (yellow), and γH2AX (red). ( D ) Fold change of nuclear 53BP1 foci per cell relative to DMSO. Individual points in the graph represent biological replicates ( n = 3). The graph depicts the mean ± SD and statistical significance was assessed by one sample t test and indicated by numerical P -values. ( E ) Fold change of nuclear γH2AX foci per cell relative to DMSO otherwise as in panel (D).

Article Snippet: Human osteosarcoma U2OS cells (#HTB-96), HCC cell lines PLC/PRF/5 (#CRL-8024), SNU-449 (#CRL-2234), and normal liver epithelial cells THLE-3 (#CRL-3583) were purchased from the American Type Culture Collection.

Techniques: Activity Assay, Control, Staining

Journal: Aging Cell

Article Title: Overactivation of Cdc42 GTPase Impairs the Cytotoxic Function of NK Cells From Old Individuals Towards Senescent Fibroblasts

doi: 10.1111/acel.70398

Figure Lengend Snippet: Natural killer cells from old adults reveal reduced cytotoxicity towards senescent fibroblasts. (A) Graphical illustration of the NK cell mediated target cell cytotoxicity assay. Target cells were first stained with calcein acetomethoxymethyl (AM), a vital fluorescent dye. Calcein AM is a non‐fluorescent compound that pass the intact cell membrane into the cytoplasm. Hydrolysis of calcein AM by intracellular esterases in live cells generates calcein, a hydrophilic, intensely fluorescent molecule which reliably stays in the cytoplasm. The stained target cells were next co‐cultured with NK cells isolated from young or old human or mice. NK cells exert their cytotoxicity towards target cells through the release of perforin and granzyme B. Upon lysis of target cells, the calcein dye is released and the loss of the dye is measured as a shift in fluorescence intensity by flow cytometry. Dead cells will appear to the left of the histogram, while alive cells on the right side. The percentage of dead cells can then simply be calculated and presented. (B) Graphical scheme depicts the experimental groups: Co‐cultures of NK cells from young adults with senescent human dermal fibroblasts in the top row and NK cells from old adults with senescent HDF in the bottom row. (C) Histogram (bi‐exponential scale) showing cytotoxicity of NK cells from young and old adults on different senescent HDF. RS, replicative senescent HDF, DIS, doxorubicin induced senescent HDF, IR, ionizing radiation induced senescence, CA, chronologically aged HDF (~75 years). The peak in the left part of the histogram showing the dead cell population and the percentage of dead cells. (D) The graph depicts the percentage of senescent HDF death ( y ‐axis) by NK cells isolated from young and old human adult. Data were represented as mean (Percentage of senescent fibroblast death) ± SEM. N = 5. Two tailed Student's t ‐test was used to assess the significance between young and old groups for each of senescence model. (E) Illustration of the experimental design showing cytotoxic activity of NK cells derived from bone marrow and spleen of young and old mice against aged murine dermal fibroblasts (MDF). (F) Histogram depicting cytotoxicity of NK cells from young and old mice on old MDF. The peak in the left part of the histogram showing the dead cell population and the percentage of dead cells. (G) The graph depicts the percentage of senescent MDF death ( y ‐axis) by NK cells isolated from young and old mice. Data were represented as mean (Percentage of senescent fibroblast death) ± SEM. N = 3. Each mouse NK cell sample used in the cytotoxicity assay was the pool of NK cells isolated from three different mice. One‐way ANOVA, followed by Bonferroni multiple comparison test was used to find the significance among the groups.

Article Snippet: Human dermal fibroblast , ATCC , Cat. #ATCC‐PCS‐201‐010.

Techniques: Cytotoxicity Assay, Staining, Membrane, Cell Culture, Isolation, Lysis, Fluorescence, Flow Cytometry, Two Tailed Test, Activity Assay, Derivative Assay, Comparison

Journal: Aging Cell

Article Title: Overactivation of Cdc42 GTPase Impairs the Cytotoxic Function of NK Cells From Old Individuals Towards Senescent Fibroblasts

doi: 10.1111/acel.70398

Figure Lengend Snippet: CASIN restores impairment of conjugation, degranulation and mitochondrial ATP generation in old Natural killer cells. (A) Synapse formation with conjugation of NK cells with the target senescent fibroblasts and the tubulin network pulling the perforin and granzyme B containing vesicles in the direction of the synapse. (B) Fusion of the NK cell derived secretory granules with the presynaptic membrane of NK cells and concomitant exposure of CD107a at the cell membrane and the release of perforin and granzyme B into the synaptic cleft towards the target cell. (C & D) Percentage of NK cell conjugation with senescent HDF when co‐cultured for (C) 60 and (D) 90 min at an effector to target (E:T) cell ratio of 1:1. Data were represented as mean (percentage of cell conjugation) ± SEM, N = 3. (E) Degranulation of NK cells when co‐cultured with senescent HDF for 7 h at an effector to target (E:T) cell ratio of 10:1. Data were represented as mean (mean fluorescence intensity) ± SEM, N = 6. One‐way ANOVA, followed by Bonferroni multiple comparison test was used to find the significance among the groups in C, D and E. (F) Seahorse flux analysis showed quantification of ATP generated by NK cells treated with vehicle from young donors and from NK cells treated with either vehicle or CASIN from old donors. The ATP generation either by glycolysis or by oxidative phosphorylation and total ATP was assessed. Data were represented as mean (ATP level) ± SEM, N = 7. Two‐way ANOVA, followed by Bonferroni multiple comparison test was used to find the significance among the groups. (G) Mitochondrial structure showing the chemical structure of the mitochondrial fluorescent probe JC‐1 that can form J‐aggregates (red) and J‐monomers (green) indicating high and low mitochondrial membrane potential, respectively. (H) Flow cytometry analysis of J‐aggregates (red) and J‐monomers (green) of young NK cells treated with vehicle, old NK cells treated with either vehicle or CASIN. (I) The graph depicts the percentage of J‐aggregates (Q2 population of figure H) of young NK treated with vehicle, and old NK cells treated with either vehicle or CASIN. Data were represented as mean (percentage of cell with J‐aggregate) ± SEM, N = 6. (J) Quantification of the ratio of J‐aggregates to J‐monomers from young and old NK treated with vehicle and old NK cells treated with CASIN. Data were represented as mean (ratio) ± SEM, N = 5. One‐way ANOVA, followed by Bonferroni multiple comparison test was used to find the significance among the groups in I and J.

Article Snippet: Human dermal fibroblast , ATCC , Cat. #ATCC‐PCS‐201‐010.

Techniques: Conjugation Assay, Derivative Assay, Membrane, Cell Culture, Fluorescence, Comparison, Generated, Phospho-proteomics, Flow Cytometry

Journal: Aging Cell

Article Title: Overactivation of Cdc42 GTPase Impairs the Cytotoxic Function of NK Cells From Old Individuals Towards Senescent Fibroblasts

doi: 10.1111/acel.70398

Figure Lengend Snippet: CASIN treatment improves the cytotoxic ability of Natural killer cells from old humans and mice. (A) Graphical illustration of experimental plan, where young NK cells treated with vehicle and old NK cells treated with either vehicle or CASIN for 8 h and thereafter subjected to co‐culture with target senescent HDF exerting their differential killing ability. (B) Representative histograms depicting the killing ability of different experimental groups as measured by flow cytometry. Peak at the left side of histogram, showing the dead senescent HDF population with percentage of dead cells. (C) Quantification of the percentage of target senescent HDF death executed by young NK cells treated with vehicle, and old NK cells treated with either vehicle or CASIN. Data were represented as mean (Percentage of senescent fibroblast death) ± SEM. N = 4. (D) Representative histograms show the distribution of K562 killing by young NK cells treated with vehicle, and old NK cells treated with either vehicle or CASIN. Peak at the left side of histogram, showing the dead K562 population with percentage of dead cells. (E) Graph shows the percentage of target cell (K562) death mediated either by young NK cells treated with vehicle or by old NK cells treated with either vehicle or CASIN. Data were represented as mean (Percentage of K562 lysis) ± SEM. N = 4. One‐way ANOVA, followed by Bonferroni multiple comparison test was used to find the significance among the groups in C and E. (F) Illustration of the experimental design for treatment of young mice (average age 120 days) treated with vehicle and old mice (average age 650 days) treated with either vehicle or CASIN. Following treatment, NK cells were isolated from spleen and bone marrow and subjected to co‐cultures with murine dermal fibroblasts (MDF) derived from old mice (average age 650 days). (G) Flow cytometry with representative histograms depicting old/senescent MDF killing by NK cells isolated from bone marrow (left panel) and spleen (right panel) of vehicle and CASIN treated old mice. Peak at the left side of histogram, showing the dead old MDF population with percentage of dead cells. (H) Quantification of the percentage of old/senescent MDF killing by NK cells isolated from bone marrow and spleen of young and old mice treated with vehicle and old mice treated with CASIN. Data were represented as mean (percentage of old/senescent MDF lysis) ± SEM, N = 4, where each group contains pool of NK cells isolated from 4 different mice of same treatment group. Two‐way ANOVA, followed by Bonferroni multiple comparison test was used to find the significance among the groups. (I) Graphical summary. Unrestrained Cdc42 activity causes failure of old NK cells to kill senescent fibroblasts. Unrestrained Cdc42 activity disrupts the microtubular network and impaired mitochondrial ATP resulting in reduced conjugation, and impaired degranulation of lytic vesicles into the synaptic cleft with reduced cytotoxicity. CASIN can attenuate all these steps and in part attenuate the killing of senescent fibroblasts (senescent HDF).

Article Snippet: Human dermal fibroblast , ATCC , Cat. #ATCC‐PCS‐201‐010.

Techniques: Co-Culture Assay, Flow Cytometry, Lysis, Comparison, Isolation, Derivative Assay, Activity Assay, Conjugation Assay