Journal: Nature Communications

Article Title: PTEN expression by an oncolytic herpesvirus directs T-cell mediated tumor clearance

doi: 10.1038/s41467-018-07344-1

Figure Lengend Snippet: HSV-P10 construction and characterization. a Graphical representation of PTEN and PTENα coding sequences showing the mutations engineered in PTENα start codons to enhance translation of full-length PTENα. b – c Graphical representation of the DNA structure of wild-type F-strain HSV1 backbone showing doubly deleted γ34.5 genes, and gene-disrupting insertional ICP6 locus containing an enhanced green fluorescent protein (eGFP) gene with PTENα insertion (HSV-P10) or without PTENα (HSVQ). d PTENα production in infected tumor cell lysates over time. The indicated tumor cells were infected with HSV-P10 at MOI = 0.5 and harvested 0–12 hpi and probed for PTEN by western blot

Article Snippet: Primary (from CST: PTEN (9559), Akt (4691), pAkt-S473 (9271), from abcam: HSV1 ICP4 (ab6514), GAPDH (ab9484)) and secondary antibodies (from abcam: anti-mouse HRP (ab6789), anti-rabbit HRP (ab6721) can be found in Supplementary Data .

Techniques: Infection, Western Blot

Journal: Nature Communications

Article Title: PTEN expression by an oncolytic herpesvirus directs T-cell mediated tumor clearance

doi: 10.1038/s41467-018-07344-1

Figure Lengend Snippet: HSV-P10 induces enhanced immune cell influx towards infected tumors. Tumor-bearing FVB/N mice were treated with 1e5 pfu of HSVQ, HSV-P10, or saline control 8 days post tumor implantation. a Stitched 4X bright field images of H&E stained sagittal sections of mouse brains 3 days post virus injection. b Representative 20X bright field images of H&E, Keratin 8, F4/80, NKp46, CD3, CD8α, PD-L1, and HSV1 (GFP) from sagittal sections of mouse brains 3 days post virus injection. Scale = 0.1 mm. c Ratio of macrophages (CD11b + F4/80 + CD45 bright ) to microglia (CD11b + F4/80 + CD45 dim ). d Percentage of cells in c positive for MHC-II. e Dendritic cell (CD11c + CD80 + MHC-II + ) infiltration. f NK cell (CD49b + NKp46 + ) infiltration. g CD8 + T-cell (CD3 + CD4 + ) infiltration. h CD4 + T-cell (CD3 + CD8 + ) infiltration. Data shown are averages ± s.d ( n = 3/group) Statistical significance was assessed by one-way ANAOVA ( n = 3, * p < 0.05, *** p < 0.001, **** p < 0.0001). Gating strategies for ( c – h ) are decribed in supplementary figure

Article Snippet: Primary (from CST: PTEN (9559), Akt (4691), pAkt-S473 (9271), from abcam: HSV1 ICP4 (ab6514), GAPDH (ab9484)) and secondary antibodies (from abcam: anti-mouse HRP (ab6789), anti-rabbit HRP (ab6721) can be found in Supplementary Data .

Techniques: Infection, Tumor Implantation, Staining, Injection

Journal: Advanced Science

Article Title: Polyplex Nanomicelle‐Mediated Pgc‐1α4 mRNA Delivery Via Hydrodynamic Limb Vein Injection Enhances Damage Resistance in Duchenne Muscular Dystrophy Mice

doi: 10.1002/advs.202409065

Figure Lengend Snippet: Expression levels of PGC‐1α and associated genes in dystrophic muscles. A) Western blotting of PGC‐1α1 and PGC‐1α4 protein levels ( n = 3 mice). B) Quantitative analysis of the western blot bands normalized to GAPDH. C) Relative mRNA expression levels of total Pgc‐1α (Pgc‐1α1,2,3, and 4), Pgc‐1α1, and Pgc‐1α4. D–K) Relative mRNA levels of genes associated with PGC‐1α in various aspects including muscle hypertrophy (D), myogenic induction (E), mitochondrial activity (F), angiogenesis (G), neuromuscular junction (NMJ) (H), myofiber type (I), metabolism (J), and inflammation (K). All analyses were performed using plantar flexor muscles. Data are presented as mean ± SEM ( n = 6 mice), and Welch's t ‐test was used ( *** p < 0.001, ** p < 0.01, and * p < 0.05).

Article Snippet: Membranes were blocked with 5% non‐fat dry milk and incubated with primary antibodies against PGC‐1α (sc‐518025, Santa Cruz; ST1202, Calbiochem) and loading controls GAPDH (D16H11, Cell Signaling Technology) and α‐tubulin (T6199, Sigma–Aldrich).

Techniques: Expressing, Muscles, Western Blot, Activity Assay

Journal: Advanced Science

Article Title: Polyplex Nanomicelle‐Mediated Pgc‐1α4 mRNA Delivery Via Hydrodynamic Limb Vein Injection Enhances Damage Resistance in Duchenne Muscular Dystrophy Mice

doi: 10.1002/advs.202409065

Figure Lengend Snippet: General upregulation of dysregulated genes after nanomicelle‐delivered Pgc‐1α4 mRNA treatment. A,B) Western blotting images of PGC‐1α4 protein expression on day 2 (A) and day 7 (B). C) Quantification of western blotting bands normalized to GAPDH (for day 2, n = 5 for the LNP + α4 group, n = 6 for all other groups; for day 7, n = 4 for the LNP + α4 group, n = 5 for the micelle + Fluc group, and n = 6 for the micelle + α4 group). D–L) Relative mRNA levels of genes associated with PGC‐1α on day 2 and day 7. All analyses were performed using plantar flexor muscle samples. Data are presented as mean ± SEM ( n = 6 mice), and one‐way ANOVA followed by Tukey's post hoc test was used ( *** p < 0.001, ** p < 0.01, and * p < 0.05).

Article Snippet: Membranes were blocked with 5% non‐fat dry milk and incubated with primary antibodies against PGC‐1α (sc‐518025, Santa Cruz; ST1202, Calbiochem) and loading controls GAPDH (D16H11, Cell Signaling Technology) and α‐tubulin (T6199, Sigma–Aldrich).

Techniques: Western Blot, Expressing