lats  (Proteintech)

Journal: bioRxiv

Article Title: aPKC-ζ III promotes trophoblast fusion by altering Par-3 interactions with Hippo Signaling Kinase LATS1

doi: 10.64898/2025.12.05.692666

Figure Lengend Snippet: Hippo signaling is altered by aPKC-ζ III in trophoblasts (A) Representative XY-plane (top) and Z-projection (bottom) images of 9-12 week placenta explants 48 hours post ST-denudation and treated with PRKCZ -siRNA, LATS inhibitor, or a both stained for E-cad (E-cadherin; green), phalloidin (magenta), and nuclei (blue). (B) Summary data of relative ST coverage; Krusal-Wallis test with Dunn’s multiple comparisons; *p ≤ 0.05, **p ≤ 0.01; n=6. (C) Graphical depiction of hypothesized interactions between aPKC-ζ III and Par-3 resulting in phosphorylation of YAP. (D) Western blotting of EGFP immunoprecipitation of Par-3-EGFP with LATS1-Myc +/-aPKC-ζ III-FLAG. (E) Summary data of relative LATS1 immunoprecipitated with Par-3; Kruskal-Wallis test with Dunn’s multiple comparisons test; *p ≤ 0.05; n=3. (F and H) Representative western blot of (F) p-YAP (phospho-Ser127) and total YAP signal or (H) p-LATS (phospho-Ser909) and total LATS from aPKC-ζ III KO cells at T=0 (Control) or T=2 after Br-cAMP +/- aPKC-ζ III reintroduction. (G and I) Summary data of relative (G) p-YAP/total YAP and II) p-LATS1/total LATS1; Kruskal-Wallis test with Dunn’s multiple comparisons test; *p ≤ 0.05, **p ≤ 0.01; n=4-5. All graphs show mean +/- S.E.M.

Article Snippet: After 24 hours, the tissue was vigorously washed to remove ST, debris, and treated with PRKCZ – targeting siRNA KD (Dharmacon, J-003526-14), non-targeting control siRNA (Dharmacon, D-001810-10), 5 μM myristoylated aPKC pseudosubstrate inhibitor (Invitrogen, 77749), or 3μM LATS inhibitor (TDI-011536; MedChemExpress, HY-150042).

Techniques: Staining, Phospho-proteomics, Western Blot, Immunoprecipitation, Control

Journal: Molecular Therapy. Methods & Clinical Development

Article Title: Durable tissue-specific transgene expression in newborn mice following intraperitoneal delivery of non-cytotoxic HSV vectors

doi: 10.1016/j.omtm.2025.101573

Figure Lengend Snippet: Comparison of JΔNI7 vector transgene expression cassettes in vivo (A) Schematic structure of KOS, JΔNI7-GWL1, and pENTR recombinant plasmids for construction of the viral vectors LAT-Luc 7CMV , LAT-Luc 7EF1α , LAT-Luc 7CAG , and LAT-Luc 7UbC . Open boxes indicate the terminal and internal inverted repeats. Black boxes indicate the deleted (Δ) genes ( ICP0 , ICP4 , and ICP27 ) or region (JOINT). The ICP47 promoter and translation initiation codon are included in the joint deletion. The ICP22 IE gene is converted to an early-expression kinetics (βICP22) by promoter TAATGARAT deletion. JΔNI7-GWL1 contains a ubiquitin C promoter (UbCp)-mCherry cassette with the SV40 polyA region in the deleted terminal ICP4 locus. JΔNI7-GWL1 contains in addition a Gateway (GW) recombination cassette in the terminal LAT locus between the LATP2 enhancer-type element and CTRL2. Luciferase expression cassettes controlled by different promoters were introduced into the JΔNI7-GWL1 vector genome by LR recombination between the GW site and each of the pENTR plasmids. (B) In vivo imaging of transgene expression in the vector-injected mice. Neonatal mice were injected i.p. at 1 × 10 10 gc/animal with PBS or vectors LAT-Luc 7CMV (CMV promoter), LAT-Luc 7EF1α (EF1α promoter), LAT-Luc 7CAG (CAG promoter), or LAT-Luc 7UbC (UbC promoter). Bioluminescence was imaged at the dorsal and ventral sides at 1, 2, 3, and 4 weeks psi. (C and D) Quantification of bioluminescence in vector-injected mice measured at the dorsal (C) and ventral (D) sides at 1, 2, 3, and 4 weeks psi (n = 7–11/group). (E) Persistence of transgene expression in vector-injected mice. Neonatal mice were injected i.p. with PBS or 1 × 10 10 gc LAT-Luc 7CAG . Bioluminescence was imaged at 20 and 24 weeks psi. (F) Quantification of bioluminescence in vector-injected mice measured at the dorsal and ventral sides at 1, 2, 4, 8, 12, 16, 20, and 24 weeks psi ( n = 6/group). Statistical significance in all experiments was evaluated with one-way ANOVA and post hoc Dunnett’s (C, D) or Tukey’s (F) multiple comparison tests. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. Data are reported as means (SD) (C, D) or means (SE) (F).

Article Snippet: For the construction of LAT-Luc 7CMV , LAT-Luc 7EF1α , LAT-Luc 7CAG , and LAT-Luc 7UbC HSV-BAC DNA, in vitro LR Clonase (Invitrogen) reactions were performed to recombine JΔNI7-GWL1 BAC DNA with pENTR-CMV-RFLuc, pENTR-EF1α-RFLuc, pENTR-CAG-RFLuc, and pENTR-UbC-RFLuc respectively.

Techniques: Comparison, Plasmid Preparation, Expressing, In Vivo, Recombinant, Ubiquitin Proteomics, Luciferase, In Vivo Imaging, Injection