Pegfp c3 lats1

Manufactured by Addgene

PEGFP C3-Lats1 is a plasmid vector that contains the gene for the Lats1 protein fused to the green fluorescent protein (GFP). Lats1 is a serine/threonine-protein kinase that plays a role in the Hippo signaling pathway, which regulates cell proliferation and apoptosis.

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Lab products found in correlation

Pegfp c3 hyap1, by Addgene (2 mentions) Lipofectamine rnaimax transfection reagent, by Thermo Fisher Scientific (2 mentions) Am4611, by Thermo Fisher Scientific (1 mentions) Pegfp c3 mst2, by Addgene (1 mentions) Lipofectamine 3000 transfection reagent, by Addgene (1 mentions) Pgfp c3 vinculin, by Addgene (1 mentions) Cmv gfp nmhc 2 b, by Addgene (1 mentions) Pcdna3 p53 wt, by Addgene (1 mentions) Prl tk, by Promega (1 mentions)

3 protocols using pegfp c3 lats1

Plasmid Construction and Characterization

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pEGFP-C3-hYAP1 and pEGFP C3-Lats1 (Addgene plasmid # 19053) were gifts from Marius Sudol (Addgene plasmid # 17843). HA-AMOT p130 was a gift from Kunliang Guan (Addgene plasmid # 32821) pcDNA3.1-ccdB-Nanoluc was a gift from Mikko Taipale (Addgene plasmid # 87067). GFP-ASPP2 was a gift from Charles D. Lopez.

Hennigan R.F., Thomson C.S., Stachowski K., Nassar N, & Ratner N. (2023). Merlin tumor suppressor function is regulated by PIP2-mediated dimerization. PLOS ONE, 18(2), e0281876.

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Overexpression and RNAi of YAP/TEAD Pathway

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For overexpression experiments, YAP–HaloTag CRISPR knock-in U-2 OS cells were transfected with pEGFP-C3-Lats1 (Addgene plasmid # 19053) or pEGFP C3-Mst2 (Addgene plasmid # 19056), both gifts from Marius Sudol, pRK5-Myc-Fus-R495X,⁶⁸ (link) gift from Jiou Wang, or EGFP-TAZ (made in the Cai lab) using Lipofectamine 3000 Transfection Reagent (cat. no. L3000015), for 16 h. For RNAi experiments, a mixture of Lats1 siRNA (Thermo Fisher, Silencer Select s17393) and Lats2 siRNA (Thermo Fisher, Silencer Select s25503) was used at a final concentration of 10 nM, or the scrambled negative control siRNA was used at a final concentration of 20 nM (Thermo Fisher, AM4611), and were transfected into cells using the Lipofectamine RNAiMAX transfection reagent (Thermo Fisher, cat. no. 13778075) for 48 h. For siTEAD1 experiments, the TEAD1 siRNA (Thermo Fisher, Silencer Select s13962) or scrambled negative control siRNA was used at a final concentration of 10 nM (Thermo Fisher, AM4611), and were transfected into cells using the Lipofectamine RNAiMAX transfection reagent (Thermo Fisher, cat. no. 13778075) for 48 h, after which cells were replated for live cell imaging and RT-qPCR.

Hao S., Lee Y.J., Benhamou Goldfajn N., Flores E., Liang J., Fuehrer H., Demmerle J., Lippincott-Schwartz J., Liu Z., Sukenik S, & Cai D. (2024). YAP condensates are highly organized hubs. iScience, 27(6), 109927.

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Plasmid Sourcing for Cell Biology

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mApple-N1 (Addgene plasmid no. 54567) was a gift from M. Davidson. pSK ZO-1 (Addgene plasmid no. 30316) was a gift from J. Anderson and A. Fanning, CMV-GFP-NMHC II-B (Addgene plasmid no. 11348) was a gift from R. Adelstein, pGFP(C3)-vinculin (Addgene plasmid no. 30312) was a gift from K. Hahn, pKvenus-Scrib (Addgene plasmid no. 58738) was a gift from I. Macara, and pEGFP-C3-hYAP1 and pEGFP C3-Lats1 (Addgene plasmid nos. 19053 and 17843, respectively) were gifts from M. Sudol. HA-AMOT p130 (Addgene plasmid no. 32821) was a gift from K. Guan. pcDNA3.1-ccdB-NLuc (Addgene plasmid no. 87067) was a gift from M. Taipale. GFP-afadin was a gift from Y. Takai (107 (link)). GFP-ASPP2 was a gift from C. D. Lopez (108 (link)). p4XBS2WT-Luc (Addgene plasmid no. 16593) was a gift from B. Vogelstein. p8xGTIIC-luciferase (Addgene plasmid no. 34615) was a gift from S. Piccolo. pcDNA3 p53 WT (Addgene plasmid no. 69003) was a gift from D. Meek. Plasmids containing the open reading frames for Epb4.1l3 and Rassf8 from the Human ORFeome V1 were purchased from the DNASU Plasmid Repository at the Arizona State University. pRL-TK was purchased from Promega. The Tet-inducible plasmids, pTRE3G-BI-ZsGreen and pCMV-Tet3G, were purchased from Tanaka Holdings.

Hennigan R.F., Fletcher J.S., Guard S, & Ratner N. (2019). Proximity biotinylation identifies a set of conformation-specific interactions between Merlin and cell junction proteins. Science signaling, 12(578), eaau8749.

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