Ad shnfatc4

Manufactured by Charles River Laboratories

Ad-shNFATc4 is a laboratory reagent designed for gene silencing experiments. It is an adenoviral vector that expresses short hairpin RNA (shRNA) targeting the NFATC4 gene. The core function of Ad-shNFATc4 is to facilitate the knockdown of NFATC4 expression in cell culture models.

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Lab products found in correlation

Ad nfatc1, by Charles River Laboratories (4 mentions) Ad nfatc2, by Charles River Laboratories (4 mentions) Ad shnfatc3, by Charles River Laboratories (4 mentions)

4 protocols using ad shnfatc4

Adenoviral Vector Overexpression of NFATs

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Constructions of NFATc1/c2/c4 overexpression adenoviral vectors were prepared as previously described^{17 (link)}. The gene accession numbers of overexpressing NFATc1/c2/c4 are NM_172390, NM_173091, and NM_004554, respectively. Constructions of NFATc3/c4 short hairpin RNA (shRNA) adenoviral vectors were prepared as previously described^{17 (link)}. These overexpression adenoviral vectors containing Ad-NFATc1, Ad-NFATc2, Ad-shNFATc3 and Ad-shNFATc4 were obtained from Vigenebio. To confirm the role of NFATc3 or NFAtc4 in myoblast cells, Ad-shCtrl, Ad-shNFATc3, or Ad-shNFATc4 (1 × 10⁹ pfu) was added to the corresponding culture dishes one day before ISO treatment. Then, these cells were replaced with differentiation medium for further observation.

Yue J., Xu W., Xiang L., Chen S.J., Li X.Y., Yang Q., Zhang R.N., Bao X., Wang Y., Mbadhi M., Liu Y., Yao L.Y., Chen L., Zhao X.Y., Hu C.Q., Zhang J.X., Zheng H.T., Wu Y., Chen S.Y., Li S., Lv J., Shi L.L, & Tang J.M. (2023). Continuous exposure to isoprenaline reduced myotube size by delaying myoblast differentiation and fusion through the NFAT-MEF2C signaling pathway. Scientific Reports, 13, 436.

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Adenoviral Vectors for Myoblast Regulation

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Ezrin-, L-periaxin-, and NFATc1/c2-overexpressing adenoviral vectors were prepared as previously described [15 (link)]. The gene accession numbers of overexpressing-Ezrin, L-periaxin, and NFATc1/c2 are NM_172390 and NM_173091, respectively. The adenoviral vectors carrying short hairpin RNA (shRNA) for knockdown of Ezrin, L-periaxin and NFATc3/c4 were prepared as previously described (Hicks et al., 2014). These overexpression adenoviral vectors containing Ad-NFATc1, Ad-NFATc2, Ad-shNFATc3 and Ad-shNFATc4 were obtained from Vigenebio. To confirm the role of L-periaxin in myoblasts, Ad-Null, Ad-Periaxin, or Ad-shPeriaxin (1 × 10⁹ pfu) was added to the corresponding culture dishes one day before Ad-Ezrin or Ad-shEzrin was added. To confirm the role of NFATc3 or NFAtc4 in myoblasts, Ad-Null, Ad-shNFATc3, or Ad-shNFATc4 (1 × 10⁹ pfu) was added to the corresponding culture dish one day before Ad-Ezrin or Ad-shEzrin was added. Then, the proliferation medium was replaced with differentiation medium for further observation. The successful knockdown and overexpression of exogenous genes was measured by detecting the His-tag, Ezrin and L-periaxin (Additional file 1: Figure S2–S3).

Zhang R.N., Bao X., Liu Y., Wang Y., Li X.Y., Tan G., Mbadhi M.N., Xu W., Yang Q., Yao L.Y., Chen L., Zhao X.Y., Hu C.Q., Zhang J.X., Zheng H.T., Wu Y., Li S., Chen S.J., Chen S.Y., Lv J., Shi L.L, & Tang J.M. (2023). The spatiotemporal matching pattern of Ezrin/Periaxin involved in myoblast differentiation and fusion and Charcot-Marie-Tooth disease-associated muscle atrophy. Journal of Translational Medicine, 21, 173.

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Modulation of NFAT Transcription Factors in Myoblasts

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Construction of NFATc1/c2 overexpression adenoviral vector were prepared as previously described 15 .
The gene accession number of overexpressing-NFATc1/c2 is NM_172390 and NM_173091, respectively. Construction of NFATc3/c4 short hairpin RNA (shRNA) adenoviral vector were prepared as previously described 15 . These overexpression adenoviral vectors containing Ad-NFATc1, Ad-NFATc2, Ad-shNFATc3 and Ad-shNFATc4were obtained from Vigenebio.To con rm the role of NFATc3 or NFAtc4 on myoblast cells, the addition of Ad-shCtrl, Ad-shNFATc3, or Ad-shNFATc4 (1 × 10 9 pfu) into correspondingculture dish one day before the ISO was performed. And then these cells were replaced with differentiation medium for further observation.

Zhang R., Wang Y., Liu Y., Bao X., Xu W., Yao L., Mbadhi M.N., Chen L., Hu C., Zheng H., Zhang Z., Zhang J., Wu Y., Lv Y., Chen S., Li S., Chen S., Ren Z., Shi L., & Tang J. (2020). Ezrin regulated myoblast differentiation/fusion and muscle fiber specialization through PKA-NFAT-MyoD/MEF2csignalling pathway.

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Adenoviral Regulation of Myoblast

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Construction of Ezrin, L-Periaxin, NFATc1/c2 overexpression adenoviral vector were prepared as previously described 15 . Constructions of Ezrin, L-Periaxin and NFATc3/c4 short hairpin RNA (shRNA) adenoviral vector were prepared as previously described 15 . These overexpression adenoviral vectors containing Ad-NFATc1, Ad-NFATc2, Ad-shNFATc3 and Ad-shNFATc4 were obtained from Vigenebio. To con rm the role of L-Periaxin in myoblast, the addition of Ad-Null, Ad-Periaxin, or Ad-shPeriaxin (1×10 9 pfu) into corresponding culture dish one day before Ad-Ezrin or Ad-shEzrin was added. To con rm the role of NFATc3 or NFAtc4 on myoblast, the addition of Ad-Null, Ad-shNFATc3, or Ad-shNFATc4 (1×10 9 pfu) into corresponding culture dish one day before Ad-Ezrin or Ad-shEzrin was added.
And then proliferation medium was replaced with differentiation medium for further observation.

Zhang R., Bao X., Wang Y., Li X., Mbadhi M.N., Liu Y., Xu W., Yao L., Chen L., Zhao X., Hu C., Zhang J., Zheng H., Wu Y., Chen S., Li S., Chen S., Lv J., Shi L., & Tang J. (2021). Ezrin Prompted Myoblast Differentiation and Muscle Fiber Specialization and Gastrocnemius Muscle Repair in Peroneal Nerve Injury Model through PKA-NFATs Signaling Pathway.

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