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