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Plpcx cx43 ires gfp

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The PLPCX-Cx43-IRES-GFP is a plasmid that contains the coding sequence for the connexin 43 (Cx43) protein and the green fluorescent protein (GFP) under the control of an internal ribosome entry site (IRES) element. The core function of this plasmid is to express Cx43 and GFP in transfected cells.

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Lab products found in correlation

Cx46 cdna, by R&D Systems (2 mentions) Pcdna3.1 hygro gjc1 cx45 cloneid, by GenScript (2 mentions) Pcdna3.1 hygro gja4 cx37 cloneid, by GenScript (2 mentions) Pbabepuro gfp lc3, by Addgene (1 mentions) Fuw mcherry gfp lc3, by Addgene (1 mentions) Lipofectamine 3000, by Thermo Fisher Scientific (1 mentions) Geneticin g418, by Thermo Fisher Scientific (1 mentions)

3 protocols using plpcx cx43 ires gfp

1

Establishing Stable Cell Lines with Fluorescently Tagged Proteins

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The following plasmids were obtained from Addgene: pBABEpuro GFP‐LC3 from Jayanta Debnath (Addgene plasmid #22405)[16] and FUW mCherry‐GFP‐LC3 from Anne Brunet (Addgene plasmid #110060).[17] Cx43‐mCherry was constructed on pLPCX‐Cx43‐IRES‐GFP from Trond Aasen (Addgene plasmid #65433).[18] Transfection of HeLa, PC12, and N2A cells with indicated plasmids was performed with Lipofectamine 3000 (Invitrogen) according to the manufacturer's instructions. Transduced cells were grown in the medium as described above. Selection of transduced cells was performed with 1 mg mL−1 Geneticin (G418) (Thermo Fisher Scientific) for about 15 days. High‐expressing cell clones were identified using fluorescence microscopy. Following selection, stably transfected HeLa and PC12 cells were cultured and supplemented with 500 µg mL−1 G418, and N2A cells were cultured with 800 µg mL−1 G418.
Li B., Wei Y., Li Q., Chen N., Li J., Liu L., Zhang J., Wang Y., Sun Y., Shi J., Wang L., Shao Z., Hu J, & Fan C. (2021). Nanomechanical Induction of Autophagy‐Related Fluorescence in Single Cells with Atomic Force Microscopy. Advanced Science, 8(24), 2102989.
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2

Engineered Connexin Expression Constructs

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The Cx46 expression vector was created by inserting the Cx46 cDNA (catalog# RDC0535, R&D Systems) between the HindIII and XbaI sites of pEGFP-N3, excising the GFP tag. This backbone was used for site-directed mutagenesis to introduce the L11S, T19M, and cysless mutations, using the primers shown in Table S2. The primers for the cysless mutant were designed so that the PCR reactions must be performed sequentially from the N-terminus to the C-terminus.
pLPCX-Cx43-IRES-GFP was obtained from Addgene (#65433). pcDNA3.1/Hygro(+)-GJC1 (Cx45; cloneID: OHu04829) and pcDNA3.1/Hygro(+)-GJA4 (Cx37; cloneID: OHu33346) were obtained from GenScript.
Mulkearns-Hubert E.E., Torre-Healy L.A., Silver D.J., Eurich J.T., Bayik D., Serbinowski E., Hitomi M., Zhou J., Przychodzen B., Zhang R., Sprowls S.A., Hale J.S., Alban T.J., Berezovsky A., Bell B.A., Lockman P.R., Jha B.K, & Lathia J.D. (2019). Development of a Cx46 Targeting Strategy for Cancer Stem Cells. Cell reports, 27(4), 1062-1072.e5.
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3

Engineered Connexin Expression Constructs

Check if the same lab product or an alternative is used in the 5 most similar protocols
The Cx46 expression vector was created by inserting the Cx46 cDNA (catalog# RDC0535, R&D Systems) between the HindIII and XbaI sites of pEGFP-N3, excising the GFP tag. This backbone was used for site-directed mutagenesis to introduce the L11S, T19M, and cysless mutations, using the primers shown in Table S2. The primers for the cysless mutant were designed so that the PCR reactions must be performed sequentially from the N-terminus to the C-terminus.
pLPCX-Cx43-IRES-GFP was obtained from Addgene (#65433). pcDNA3.1/Hygro(+)-GJC1 (Cx45; cloneID: OHu04829) and pcDNA3.1/Hygro(+)-GJA4 (Cx37; cloneID: OHu33346) were obtained from GenScript.
Mulkearns-Hubert E.E., Torre-Healy L.A., Silver D.J., Eurich J.T., Bayik D., Serbinowski E., Hitomi M., Zhou J., Przychodzen B., Zhang R., Sprowls S.A., Hale J.S., Alban T.J., Berezovsky A., Bell B.A., Lockman P.R., Jha B.K, & Lathia J.D. (2019). Development of a Cx46 Targeting Strategy for Cancer Stem Cells. Cell reports, 27(4), 1062-1072.e5.
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