Cmv gfp nmhc 2 b

Manufactured by Addgene

CMV-GFP-NMHC II-B is a plasmid that expresses non-muscle myosin heavy chain IIB (NMHC II-B) fused with green fluorescent protein (GFP) under the control of the cytomegalovirus (CMV) promoter.

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Lab products found in correlation

Cmv gfp nmhc 2 a, by Addgene (2 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions) Px335 u6 chimeric bb cbh hspcas9n d10a, by Addgene (1 mentions) Pspcas9 bb 2a puro px459 v2, by Addgene (1 mentions) Geneart, by Thermo Fisher Scientific (1 mentions) Pgfp c3 vinculin, by Addgene (1 mentions) Pcdna3 p53 wt, by Addgene (1 mentions) Pegfp c3 hyap1, by Addgene (1 mentions) Pegfp c3 lats1, by Addgene (1 mentions) Prl tk, by Promega (1 mentions)

3 protocols using cmv gfp nmhc 2 b

Plasmid Transfections and CRISPR Modifications

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Transfections were carried out using Lipofectamine 2000 (ThermoFisher Scientific #11668027) according to manufacturer’s instructions. The cells were transfected 48 hr prior to the experiment. CMV-GFP-NMHC IIA (Addgene #11347) and CMV-GFP-NMHC IIB (Addgene #11348) were gifts from Robert Adelstein (Wei and Adelstein, 2000 (link)). NMHC IIB-mApple was a gift from Michael Davidson (Addgene #54931). pSPCas9(BB)−2A-Puro (PX459) V2.0 (Addgene #62988) and pX335-U6-Chimeric_BB-CBh-hSpCas9n(D10A) (Addgene #42335) were gifts from Feng Zhang. Guide sequences for the generation of NMHC II depleted cells or GFP knock-in cells were introduced by digesting the plasmids with BbsI and subsequent ligation (Ran et al., 2013 (link)). pMK-RQ-MYH9 and pMK-RQ-MYH10 donor plasmids for homology directed repair were constructed by flanking the coding sequence of eGFP with 800 bp homology arms upstream and downstream of the double strand break near the start codon of MYH9 or MYH10 exon two and the plasmids were synthesized by GeneArt (ThermoFisher Scientific).

Weißenbruch K., Grewe J., Hippler M., Fladung M., Tremmel M., Stricker K., Schwarz U.S, & Bastmeyer M. (2021). Distinct roles of nonmuscle myosin II isoforms for establishing tension and elasticity during cell morphodynamics. eLife, 10, e71888.

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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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Xenopus Lmo7 Expression Constructs

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Xenopus laevis Lmo7 cDNA clone was obtained from Dharmacon. Full-length and truncated forms of Lmo7 were PCR amplified and cloned into a pCS107-Flag, -HA or -GFP expression vector using standard protocols. Primers used for the amplification are in Supplemental Table 1. pCS2-RFP-HA-Wtip was previously described (Chu et al., 2018) . pCS2-GFP-NMIIA and pCS2-GFP-NMIIB were subcloned from CMV-GFP-NMHC II-A, CMV-GFP-NMHC II-B (Addgene plasmids # 11347, 11348) (Wei and Adelstein, 2000) . pCS2-GFP-a-actinin 4 was constructed from pCS2-hiActTS-GR (a gift of Tatsuo Michiue). pCS2-ROK-C and pCS2-Mypt1CA were from Florence Marlow (Marlow et al., 2002) and David Kimelman (Weiser et al., 2009) , respectively.

Matsuda M., Chu C., & Sokol S.Y. (2021). Lmo7 recruits myosin II heavy chain to induce apical constriction in Xenopus ectoderm.

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