Journal: Aging Cell

Article Title: Role of PI3K / AKT / MAOA in glucocorticoid‐induced oxidative stress and associated premature senescence of the trabecular meshwork

doi: 10.1111/acel.14452

Figure Lengend Snippet: Effect of inhibition of the PI3K‐AKT pathway on GIG progression in the GIG mouse model. (a) Schematic diagram depicting the construction of the GIG mouse model and the IOP elevation in GIG mice (red asterisk: Vehicle vs. DEX‐Ace; n = 10 mouse eyes). (b) Protein expression levels of pan‐AKT, p‐AKT‐S473, and p‐AKT‐T308 in TM of vehicle and DEX‐Ace groups ( n = 3 mouse eyes). (c) Protein expression levels of pan‐AKT, p‐AKT‐S473, and p‐AKT‐T308 in TM of vehicle, DEX‐Ace, LY294002 + vehicle, and LY294002 + DEX‐Ace groups ( n = 3 mouse eyes). (d) LY294002 (PI3K inhibitor; 50 mg/kg, once weekly for 8 weeks) reduced IOP in the GIG mouse model (red asterisk: Vehicle vs. DEX‐Ace; blue asterisk: DEX‐Ace vs. LY294002 + DEX‐Ace; n = 10 mouse eyes). (e) Representative images of mitochondrial morphology in the TM region from each group captured by electron microscopy. White arrows: Damaged mitochondria in the DEX‐Ace‐treated group ( n = 3 mouse eyes). Scale bar = 10 μm (upper images)/2 μm (middle images)/1 μm (lower images). CB, ciliary body; S, Sclera; SC, Schlemm's canal; TM, Trabecular meshwork. Data are presented as mean ± SD. Unpaired t ‐test (a, b). One‐way ANOVA followed by Tukey's test (c, d). * p < 0.5, ** p < 0.01, *** p < 0.001, **** p < 0.0001. All experiments were biologically replicated at least three times.

Article Snippet: For ex vivo experiments, a GIG cell model was established using DEX (D4902, Sigma‐Aldrich, Darmstadt, Germany, 100 nM) for 72 h. LY294002 (50 μM; MedChenExpress, New Jersey, USA) or Moclobemide (50 μM; MedChenExpress, New Jersey, USA) treated the cells for 72 h. LY294002/Moclobemide and DEX were co‐added in the medium of pHTMs.

Techniques: Inhibition, Expressing, Electron Microscopy

Journal: Aging Cell

Article Title: Role of PI3K / AKT / MAOA in glucocorticoid‐induced oxidative stress and associated premature senescence of the trabecular meshwork

doi: 10.1111/acel.14452

Figure Lengend Snippet: PI3K/AKT pathway inhibition alleviated DEX‐induced oxidative stress in pHTMs. (a) LY294002 (50 μM, 72 h) inhibited DEX‐induced PI3K/AKT pathway activation in pHTMs. (b) ROS levels ( n ≥ 13 fields per group). Scale bar = 500 μm. (c) Mitochondrial superoxide levels ( n ≥ 15 fields per group). Scale bar = 200 μm. (d) PIK3R1 mRNA expression levels. (e) Protein levels of PIK3R1 and phosphorylation of AKT. (f) Representative ROS staining images ( n ≥ 14 fields per group). Scale bar = 500 μm. (g) Representative mitochondrial superoxide staining images ( n ≥ 9 fields per group). Scale bar = 200 μm. The experiments were conducted using cell strains cultured from three separate donors. Data are presented as mean ± SD. One‐way ANOVA followed by Tukey's test. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: For ex vivo experiments, a GIG cell model was established using DEX (D4902, Sigma‐Aldrich, Darmstadt, Germany, 100 nM) for 72 h. LY294002 (50 μM; MedChenExpress, New Jersey, USA) or Moclobemide (50 μM; MedChenExpress, New Jersey, USA) treated the cells for 72 h. LY294002/Moclobemide and DEX were co‐added in the medium of pHTMs.

Techniques: Inhibition, Activation Assay, Expressing, Staining, Cell Culture

Journal: Aging Cell

Article Title: Role of PI3K / AKT / MAOA in glucocorticoid‐induced oxidative stress and associated premature senescence of the trabecular meshwork

doi: 10.1111/acel.14452

Figure Lengend Snippet: Effect of inhibiting MAOA on GIG progression in the GIG mouse model. (a) Protein expression levels of MAOA in TM of vehicle, DEX‐Ace, LY294002 + vehicle, and LY294002 + DEX‐Ace groups ( n = 3 mouse eyes). (b) Effect of Moclobemide (Moc; MAOA inhibitor; 40 mg/kg, once every 2 days for 8 weeks) on IOP elevation in the GIG mouse model (red asterisk: Vehicle vs. DEX‐Ace; orange asterisk: DEX‐Ace vs. Moc + DEX‐Ace; n = 10 mouse eyes). (c) Representative images of mitochondrial morphology in the TM region from each group captured by electron microscopy. White arrows: Damaged mitochondria (n = 3 mouse eyes). Scale bar = 10 μm (upper images)/2 μm (upper images)/1 μm (lower images). CB, ciliary body; SC, Schlemm's canal; S, sclera; TM, trabecular meshwork. Data are presented as mean ± SD. One‐way ANOVA followed by Tukey's test. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. All experiments were biologically replicated at least three times.

Article Snippet: For ex vivo experiments, a GIG cell model was established using DEX (D4902, Sigma‐Aldrich, Darmstadt, Germany, 100 nM) for 72 h. LY294002 (50 μM; MedChenExpress, New Jersey, USA) or Moclobemide (50 μM; MedChenExpress, New Jersey, USA) treated the cells for 72 h. LY294002/Moclobemide and DEX were co‐added in the medium of pHTMs.

Techniques: Expressing, Electron Microscopy

Journal: Aging Cell

Article Title: Role of PI3K / AKT / MAOA in glucocorticoid‐induced oxidative stress and associated premature senescence of the trabecular meshwork

doi: 10.1111/acel.14452

Figure Lengend Snippet: PI3K/AKT/MAOA pathway modulates TM aging in the GIG mouse model. (a) KEGG analysis of DEGs between the DEX and si‐PIK3R1 + DEX groups. (b) Representative immunofluorescence images of anterior chamber angle from the vehicle and the DEX‐Ace group after 8 weeks of DEX‐Ace treatment, p21 was stained as a senescence marker, TM region was labeled using α‐smooth muscle actin (α‐SMA), and the nuclei were stained with DAPI ( n = 3 mouse eyes). Scale bar = 200 μm. (c) LY294002 (PI3K inhibitor; 50 mg/kg, once weekly for 8 weeks) inhibited the protein expression levels of aging markers p16 and p21 in TM of GIG mice ( n = 3 mouse eyes). (d) Representative immunofluorescence images. LY294002 reversed the DEX‐Ace‐induced increase of p21 expression in TM. The TM region was labeled using α‐SMA, and the nuclei were stained with DAPI. Scale bar = 200 μm. (e) Moclobemide (Moc; MAOA inhibitor; 40 mg/kg, once every 2 days for 8 weeks) inhibited the protein expression levels of aging markers p16 and p21 in TM of GIG mice ( n = 3 mouse eyes). CB, Ciliary body. Data are presented as mean ± SD. One‐way ANOVA followed by the Tukey's test. ** p < 0.01, *** p < 0.001, **** p < 0.0001. All experiments were biologically replicated at least three times.

Article Snippet: For ex vivo experiments, a GIG cell model was established using DEX (D4902, Sigma‐Aldrich, Darmstadt, Germany, 100 nM) for 72 h. LY294002 (50 μM; MedChenExpress, New Jersey, USA) or Moclobemide (50 μM; MedChenExpress, New Jersey, USA) treated the cells for 72 h. LY294002/Moclobemide and DEX were co‐added in the medium of pHTMs.

Techniques: Immunofluorescence, Staining, Marker, Labeling, Expressing

Journal: Aging Cell

Article Title: Role of PI3K / AKT / MAOA in glucocorticoid‐induced oxidative stress and associated premature senescence of the trabecular meshwork

doi: 10.1111/acel.14452

Figure Lengend Snippet: PI3K/AKT pathway inhibition prevented DEX‐induced pHTMs premature senescence. (a) Percentage of senescent cells assessed using Sa‐β‐galactosidase staining after LY294002 (50 μM, 72 h) treatment ( n ≥ 10 fields per group). Scale bar = 200 μm. (b) Cell cycle changes detected by flow cytometry with PI staining. (c) Expression of p21 and p16 at the protein level. (d) Percentage of senescent cells assessed using Sa‐β‐galactosidase staining after knocking down PIK3R1 ( n ≥ 10 fields per group). Scale bar = 200 μm. (e) Cell cycle changes detected by flow cytometry with PI staining. (f) Expression of p21 and p16 at the protein level. The experiments were conducted using cell strains cultured from three separate donors. Data are presented as mean ± SD. One‐way ANOVA followed by Tukey's test. * p < 0.05, ** p < 0.01, **** p < 0.0001.

Article Snippet: For ex vivo experiments, a GIG cell model was established using DEX (D4902, Sigma‐Aldrich, Darmstadt, Germany, 100 nM) for 72 h. LY294002 (50 μM; MedChenExpress, New Jersey, USA) or Moclobemide (50 μM; MedChenExpress, New Jersey, USA) treated the cells for 72 h. LY294002/Moclobemide and DEX were co‐added in the medium of pHTMs.

Techniques: Inhibition, Staining, Flow Cytometry, Expressing, Cell Culture

Journal: bioRxiv

Article Title: Chromatin architecture and physical constriction cooperate in phenotype switching and cancer cell dissemination

doi: 10.64898/2026.02.05.702638

Figure Lengend Snippet: a. Seurat UMAP representing the bridge integration of melanoma cell states between scRNAseq (left) and loHiC (right) obtained from a cell derived xenograft (CDX) of 12-273BM short-term patients’ culture. b. cis compartment interactions quantified in MEL and MES states identified in CDX tumors. c. Column dot plot showing the quantification of the number (left) and size (right) of TADs in MEL and MES states by loHiC. d. Genome-wide insulation score profile at TADs’ boundaries in loHiC MEL and MES states. e. TADs’ number (left) and size (right) quantified in a small cohort of FFPE MEL (n=3) and MES (n=4) tumors processed with HiC. Unpaired t-test was conducted and resulted p-values are shown. Each dot represent a patient sample.

Article Snippet: Next, FFPE samples were processed with the Arima-HiC+ FFPE Kit (Arima Genomics, #A101060) following manufacturer’s instructions.

Techniques: Derivative Assay, Genome Wide, Insulation

Journal: bioRxiv

Article Title: Chromatin architecture and physical constriction cooperate in phenotype switching and cancer cell dissemination

doi: 10.64898/2026.02.05.702638

Figure Lengend Snippet: a. Quantification of the number of CNVs as a grouped column plot across MEL, siNTC, siMS, INT and MES samples. Two-way ANOVA test was conducted. b. Column dot plot of the number of structural variants (SVs) found in the melanoma cell lines used in this study. One-way ANOVA test was performed. c. Fraction of cis -short, cis -long and trans HiC contacts in siNTC and siMS cells. Paired t-test was used. d. HiC aggregate region analysis at all TSS in siMS relative to siNTC. e. PCA plot of the most variable cCREs identified with DESeq2. f. ABC quantification of the top 500 state-specific cCRE Hubs across all cell lines. Bar plots show the average of all hubs. Standard error of the means bar are shown. Red, grey and green mark melanocytic, intermediate and mesenchymal cell lines respectively. g. Bar dot plot quantify the average methylation of 500 MES hubs in TCGA melanoma patients (n=470) classified in MEL, INT, NC (neural crest) and MES according to Tsoi . One-way ANOVA p-value summarizing NC and MES vs INT and MEL comparisons is shown. h. HiC aggregate pile up heatmap of contacts at MES hubs in FFPE patients’ specimen classified in MEL (n=3) and MES (n=4). i. Quantification of HiC contacts at the foreground of MEL and MES hubs in MEL and MES melanoma patients (n=7). Unpaired t-test p-values are shown. j. CTCF average genomic occupancy at MES hubs (n=500) in individual mesenchymal cell lines. k. Volcano plot of the differential MES cCREs activity in siCTCF MM099 cells predicted by RNAseq. In blue are marked the significantly dysregulated cCREs, in red the hubs. Y-axis dotted line show the significant p-value threshold. On top are marked the number of significantly active MES hubs (42 down, 136 up). Multiple t-test with Benjamini/Hochberg correction was used. For panels a,b,c,e,g and j, each dot is representative of a biological replicate.

Article Snippet: Next, FFPE samples were processed with the Arima-HiC+ FFPE Kit (Arima Genomics, #A101060) following manufacturer’s instructions.

Techniques: Methylation, Activity Assay, RNA sequencing