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nfat plasmid dna  (Addgene inc)


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    Addgene inc nfat plasmid dna
    Figure 1. NGF activates <t>NFAT-dependent</t> gene expression in PC12 cells. A. Schematic diagram of depicting the experimental pro- tocols. B. According to the protocol described in (A), PC12 cells transfected together with NFAT-luc (NFAT reporter, firefly luciferase) and pGL4.74[hRluc/TK] (constitutively active Renilla luciferase reporter) were co-stimulated with 20 ng/ml PMA (Phorbol 12-myristate 13-acetate, a PKC activator) and 50 mM KCl in the presence of 1 µM of BayK8644 for 5 h. After that, NFAT-dependent gene expression was determined by calculating ratios of firefly to Renilla luciferase activity. n = 7–8, ∗∗p < 0.01, Student’s t-test. C. Following the same pro- tocol described in (B), the cells were stimulated by NGF (concentration ranged from 0 ng/ml to 100 ng/ml) for 5 h. NFAT-dependent gene expression was then determined by calculating ratios of firefly to Renilla luciferase activity. n = 6–12, ∗∗∗p < 0.001 (vs 0 ng/ml NGF), One-Way ANOVA, Bonferroni post-hoc test. D. Either 0 ng/ml ((-)NGF) or 20 ng/ml NGF ((+)NGF) was applied for 5, 10, and 24 h to active PC12 cells transfected with the two luciferase reporters. NFAT-dependent gene expression was then assayed as described in (A). The ratio of NFAT-dependent gene expression in the presence of 20 ng/ml NGF to NFAT-dependent gene expression in the absence of NGF was calculated from (D), which was shown in the inset. n = 6–12. n stands for the number of well. Data are presented as mean ± S.E.M.
    Nfat Plasmid Dna, supplied by Addgene inc, used in various techniques. Bioz Stars score: 90/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/nfat plasmid dna/product/Addgene inc
    Average 90 stars, based on 15 article reviews
    nfat plasmid dna - by Bioz Stars, 2026-03
    90/100 stars

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    1) Product Images from "NGF activates NFAT via the MEK1/2 pathway in PC12 cells"

    Article Title: NGF activates NFAT via the MEK1/2 pathway in PC12 cells

    Journal: All Life

    doi: 10.1080/26895293.2022.2034670

    Figure 1. NGF activates NFAT-dependent gene expression in PC12 cells. A. Schematic diagram of depicting the experimental pro- tocols. B. According to the protocol described in (A), PC12 cells transfected together with NFAT-luc (NFAT reporter, firefly luciferase) and pGL4.74[hRluc/TK] (constitutively active Renilla luciferase reporter) were co-stimulated with 20 ng/ml PMA (Phorbol 12-myristate 13-acetate, a PKC activator) and 50 mM KCl in the presence of 1 µM of BayK8644 for 5 h. After that, NFAT-dependent gene expression was determined by calculating ratios of firefly to Renilla luciferase activity. n = 7–8, ∗∗p < 0.01, Student’s t-test. C. Following the same pro- tocol described in (B), the cells were stimulated by NGF (concentration ranged from 0 ng/ml to 100 ng/ml) for 5 h. NFAT-dependent gene expression was then determined by calculating ratios of firefly to Renilla luciferase activity. n = 6–12, ∗∗∗p < 0.001 (vs 0 ng/ml NGF), One-Way ANOVA, Bonferroni post-hoc test. D. Either 0 ng/ml ((-)NGF) or 20 ng/ml NGF ((+)NGF) was applied for 5, 10, and 24 h to active PC12 cells transfected with the two luciferase reporters. NFAT-dependent gene expression was then assayed as described in (A). The ratio of NFAT-dependent gene expression in the presence of 20 ng/ml NGF to NFAT-dependent gene expression in the absence of NGF was calculated from (D), which was shown in the inset. n = 6–12. n stands for the number of well. Data are presented as mean ± S.E.M.
    Figure Legend Snippet: Figure 1. NGF activates NFAT-dependent gene expression in PC12 cells. A. Schematic diagram of depicting the experimental pro- tocols. B. According to the protocol described in (A), PC12 cells transfected together with NFAT-luc (NFAT reporter, firefly luciferase) and pGL4.74[hRluc/TK] (constitutively active Renilla luciferase reporter) were co-stimulated with 20 ng/ml PMA (Phorbol 12-myristate 13-acetate, a PKC activator) and 50 mM KCl in the presence of 1 µM of BayK8644 for 5 h. After that, NFAT-dependent gene expression was determined by calculating ratios of firefly to Renilla luciferase activity. n = 7–8, ∗∗p < 0.01, Student’s t-test. C. Following the same pro- tocol described in (B), the cells were stimulated by NGF (concentration ranged from 0 ng/ml to 100 ng/ml) for 5 h. NFAT-dependent gene expression was then determined by calculating ratios of firefly to Renilla luciferase activity. n = 6–12, ∗∗∗p < 0.001 (vs 0 ng/ml NGF), One-Way ANOVA, Bonferroni post-hoc test. D. Either 0 ng/ml ((-)NGF) or 20 ng/ml NGF ((+)NGF) was applied for 5, 10, and 24 h to active PC12 cells transfected with the two luciferase reporters. NFAT-dependent gene expression was then assayed as described in (A). The ratio of NFAT-dependent gene expression in the presence of 20 ng/ml NGF to NFAT-dependent gene expression in the absence of NGF was calculated from (D), which was shown in the inset. n = 6–12. n stands for the number of well. Data are presented as mean ± S.E.M.

    Techniques Used: Gene Expression, Transfection, Luciferase, Activity Assay, Concentration Assay

    Figure 2. NGF recruits TrkA but does not rely on the PLC pathway to activate NFAT-dependent gene expression. A. Schematic diagram of depictingtheexperimentalprotocols.B.AdiagramdescribingthePLCpathwaystimulatedbyNGF.C.Accordingtotheprotocoldescribed in (A), NGF (20 ng/ml, 5 h stimulation) was applied and its ability to activate NFAT-dependent gene expression in the presence or absence of K252a (a TrkA inhibitor) was assayed in PC12 cells as described in Figure 1. n = 6–7, ∗∗∗p < 0.001, One-Way ANOVA, Bonferroni post-hoc test D. U73122 (a PLC inhibitor) or Ca2+-dependent phosphatase calcineurin (CaN) inhibitors (FK506, CsA) were applied to block TrkA-triggered PLC activation or PLC-IP3-Ca2+ mediated CaN activation in PC12 cells treated as explained in (A). n = 6–7 E. NGF- treated PC12 cells were co-stimulated with KCl to depolarize cells for inducing an intracellular Ca2+ increase. To verify the integrity of CaN inhibitors, the inhibitors were applied to PC12 cells prepared as described in (A) to check if they inhibit CaN which should be activated by KCl. All inhibitors were applied 30 min prior to NGF stimulation. n = 6–7, ∗∗p < 0.01, ∗∗∗p < 0.001, One-Way ANOVA, Bonferroni post-hoc test. n stands for the number of well. Data are presented as mean ± S.E.M.
    Figure Legend Snippet: Figure 2. NGF recruits TrkA but does not rely on the PLC pathway to activate NFAT-dependent gene expression. A. Schematic diagram of depictingtheexperimentalprotocols.B.AdiagramdescribingthePLCpathwaystimulatedbyNGF.C.Accordingtotheprotocoldescribed in (A), NGF (20 ng/ml, 5 h stimulation) was applied and its ability to activate NFAT-dependent gene expression in the presence or absence of K252a (a TrkA inhibitor) was assayed in PC12 cells as described in Figure 1. n = 6–7, ∗∗∗p < 0.001, One-Way ANOVA, Bonferroni post-hoc test D. U73122 (a PLC inhibitor) or Ca2+-dependent phosphatase calcineurin (CaN) inhibitors (FK506, CsA) were applied to block TrkA-triggered PLC activation or PLC-IP3-Ca2+ mediated CaN activation in PC12 cells treated as explained in (A). n = 6–7 E. NGF- treated PC12 cells were co-stimulated with KCl to depolarize cells for inducing an intracellular Ca2+ increase. To verify the integrity of CaN inhibitors, the inhibitors were applied to PC12 cells prepared as described in (A) to check if they inhibit CaN which should be activated by KCl. All inhibitors were applied 30 min prior to NGF stimulation. n = 6–7, ∗∗p < 0.01, ∗∗∗p < 0.001, One-Way ANOVA, Bonferroni post-hoc test. n stands for the number of well. Data are presented as mean ± S.E.M.

    Techniques Used: Gene Expression, Blocking Assay, Activation Assay

    Figure 3. NFAT activation by NGF depends on MEK1/2 but not PI3 K pathway. A. Schematic diagram of depicting the experimental pro- tocols. B. A diagram describing TrkA-mediated PI3 K and MEK1/2 activation by NGF. C-D. As described in Figure 2, NFAT reporter assays were performed with the exception that PI3 K inhibitor (LY294002, Wort (wortmannin) (C)) or MEK1/2 inhibitor (U0126, (D)) was applied to block TrkA-mediated PI3 K or MEK1/2 activation, respectively. All inhibitors were applied 30 min prior to NGF stimulation. n = 12–13 for (C), n = 6–12 for (D), ∗∗p < 0.01, ∗∗∗p < 0.001, One-Way ANOVA, Bonferroni post-hoc test. n stands for the number of well. Data are presented as mean ± S.E.M.
    Figure Legend Snippet: Figure 3. NFAT activation by NGF depends on MEK1/2 but not PI3 K pathway. A. Schematic diagram of depicting the experimental pro- tocols. B. A diagram describing TrkA-mediated PI3 K and MEK1/2 activation by NGF. C-D. As described in Figure 2, NFAT reporter assays were performed with the exception that PI3 K inhibitor (LY294002, Wort (wortmannin) (C)) or MEK1/2 inhibitor (U0126, (D)) was applied to block TrkA-mediated PI3 K or MEK1/2 activation, respectively. All inhibitors were applied 30 min prior to NGF stimulation. n = 12–13 for (C), n = 6–12 for (D), ∗∗p < 0.01, ∗∗∗p < 0.001, One-Way ANOVA, Bonferroni post-hoc test. n stands for the number of well. Data are presented as mean ± S.E.M.

    Techniques Used: Activation Assay, Blocking Assay

    Figure 4. NFAc3 is the predominant NFAT isoform in PC12 cells. A. After constructing standard curves for each NFAT isoform, SYBR-based qPCR was performed with total RNA isolated from PC12 cells to calculate absolute copy number of each NFAT isoform. n = 3 B. Proposed model: NFATc3, a major NFAT isoform in PC12 cells shuttles in and out of the nucleus by CaN, an NFAT phosphatase and several kinases, respectively. An increase in NFAT-dependent gene expression by NGF depends on the MEK1/2 pathway via uncharacterized mechanisms in PC12 cells. Data are presented as mean ± S.E.M.
    Figure Legend Snippet: Figure 4. NFAc3 is the predominant NFAT isoform in PC12 cells. A. After constructing standard curves for each NFAT isoform, SYBR-based qPCR was performed with total RNA isolated from PC12 cells to calculate absolute copy number of each NFAT isoform. n = 3 B. Proposed model: NFATc3, a major NFAT isoform in PC12 cells shuttles in and out of the nucleus by CaN, an NFAT phosphatase and several kinases, respectively. An increase in NFAT-dependent gene expression by NGF depends on the MEK1/2 pathway via uncharacterized mechanisms in PC12 cells. Data are presented as mean ± S.E.M.

    Techniques Used: Isolation, Gene Expression



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    Figure 1. NGF activates <t>NFAT-dependent</t> gene expression in PC12 cells. A. Schematic diagram of depicting the experimental pro- tocols. B. According to the protocol described in (A), PC12 cells transfected together with NFAT-luc (NFAT reporter, firefly luciferase) and pGL4.74[hRluc/TK] (constitutively active Renilla luciferase reporter) were co-stimulated with 20 ng/ml PMA (Phorbol 12-myristate 13-acetate, a PKC activator) and 50 mM KCl in the presence of 1 µM of BayK8644 for 5 h. After that, NFAT-dependent gene expression was determined by calculating ratios of firefly to Renilla luciferase activity. n = 7–8, ∗∗p < 0.01, Student’s t-test. C. Following the same pro- tocol described in (B), the cells were stimulated by NGF (concentration ranged from 0 ng/ml to 100 ng/ml) for 5 h. NFAT-dependent gene expression was then determined by calculating ratios of firefly to Renilla luciferase activity. n = 6–12, ∗∗∗p < 0.001 (vs 0 ng/ml NGF), One-Way ANOVA, Bonferroni post-hoc test. D. Either 0 ng/ml ((-)NGF) or 20 ng/ml NGF ((+)NGF) was applied for 5, 10, and 24 h to active PC12 cells transfected with the two luciferase reporters. NFAT-dependent gene expression was then assayed as described in (A). The ratio of NFAT-dependent gene expression in the presence of 20 ng/ml NGF to NFAT-dependent gene expression in the absence of NGF was calculated from (D), which was shown in the inset. n = 6–12. n stands for the number of well. Data are presented as mean ± S.E.M.
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    Figure 1. NGF activates <t>NFAT-dependent</t> gene expression in PC12 cells. A. Schematic diagram of depicting the experimental pro- tocols. B. According to the protocol described in (A), PC12 cells transfected together with NFAT-luc (NFAT reporter, firefly luciferase) and pGL4.74[hRluc/TK] (constitutively active Renilla luciferase reporter) were co-stimulated with 20 ng/ml PMA (Phorbol 12-myristate 13-acetate, a PKC activator) and 50 mM KCl in the presence of 1 µM of BayK8644 for 5 h. After that, NFAT-dependent gene expression was determined by calculating ratios of firefly to Renilla luciferase activity. n = 7–8, ∗∗p < 0.01, Student’s t-test. C. Following the same pro- tocol described in (B), the cells were stimulated by NGF (concentration ranged from 0 ng/ml to 100 ng/ml) for 5 h. NFAT-dependent gene expression was then determined by calculating ratios of firefly to Renilla luciferase activity. n = 6–12, ∗∗∗p < 0.001 (vs 0 ng/ml NGF), One-Way ANOVA, Bonferroni post-hoc test. D. Either 0 ng/ml ((-)NGF) or 20 ng/ml NGF ((+)NGF) was applied for 5, 10, and 24 h to active PC12 cells transfected with the two luciferase reporters. NFAT-dependent gene expression was then assayed as described in (A). The ratio of NFAT-dependent gene expression in the presence of 20 ng/ml NGF to NFAT-dependent gene expression in the absence of NGF was calculated from (D), which was shown in the inset. n = 6–12. n stands for the number of well. Data are presented as mean ± S.E.M.
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    Figure 1. NGF activates NFAT-dependent gene expression in PC12 cells. A. Schematic diagram of depicting the experimental pro- tocols. B. According to the protocol described in (A), PC12 cells transfected together with NFAT-luc (NFAT reporter, firefly luciferase) and pGL4.74[hRluc/TK] (constitutively active Renilla luciferase reporter) were co-stimulated with 20 ng/ml PMA (Phorbol 12-myristate 13-acetate, a PKC activator) and 50 mM KCl in the presence of 1 µM of BayK8644 for 5 h. After that, NFAT-dependent gene expression was determined by calculating ratios of firefly to Renilla luciferase activity. n = 7–8, ∗∗p < 0.01, Student’s t-test. C. Following the same pro- tocol described in (B), the cells were stimulated by NGF (concentration ranged from 0 ng/ml to 100 ng/ml) for 5 h. NFAT-dependent gene expression was then determined by calculating ratios of firefly to Renilla luciferase activity. n = 6–12, ∗∗∗p < 0.001 (vs 0 ng/ml NGF), One-Way ANOVA, Bonferroni post-hoc test. D. Either 0 ng/ml ((-)NGF) or 20 ng/ml NGF ((+)NGF) was applied for 5, 10, and 24 h to active PC12 cells transfected with the two luciferase reporters. NFAT-dependent gene expression was then assayed as described in (A). The ratio of NFAT-dependent gene expression in the presence of 20 ng/ml NGF to NFAT-dependent gene expression in the absence of NGF was calculated from (D), which was shown in the inset. n = 6–12. n stands for the number of well. Data are presented as mean ± S.E.M.

    Journal: All Life

    Article Title: NGF activates NFAT via the MEK1/2 pathway in PC12 cells

    doi: 10.1080/26895293.2022.2034670

    Figure Lengend Snippet: Figure 1. NGF activates NFAT-dependent gene expression in PC12 cells. A. Schematic diagram of depicting the experimental pro- tocols. B. According to the protocol described in (A), PC12 cells transfected together with NFAT-luc (NFAT reporter, firefly luciferase) and pGL4.74[hRluc/TK] (constitutively active Renilla luciferase reporter) were co-stimulated with 20 ng/ml PMA (Phorbol 12-myristate 13-acetate, a PKC activator) and 50 mM KCl in the presence of 1 µM of BayK8644 for 5 h. After that, NFAT-dependent gene expression was determined by calculating ratios of firefly to Renilla luciferase activity. n = 7–8, ∗∗p < 0.01, Student’s t-test. C. Following the same pro- tocol described in (B), the cells were stimulated by NGF (concentration ranged from 0 ng/ml to 100 ng/ml) for 5 h. NFAT-dependent gene expression was then determined by calculating ratios of firefly to Renilla luciferase activity. n = 6–12, ∗∗∗p < 0.001 (vs 0 ng/ml NGF), One-Way ANOVA, Bonferroni post-hoc test. D. Either 0 ng/ml ((-)NGF) or 20 ng/ml NGF ((+)NGF) was applied for 5, 10, and 24 h to active PC12 cells transfected with the two luciferase reporters. NFAT-dependent gene expression was then assayed as described in (A). The ratio of NFAT-dependent gene expression in the presence of 20 ng/ml NGF to NFAT-dependent gene expression in the absence of NGF was calculated from (D), which was shown in the inset. n = 6–12. n stands for the number of well. Data are presented as mean ± S.E.M.

    Article Snippet: SYBR green-based qPCR was performed with the cDNA from PC12 cells together with each NFAT plasmid DNA of known concentration (GFP-NFATc1, Addgene plasmid #11101; EGFPNFATc2, provided by Dr. Rao; EGFP-NFATc3, originally provided by Dr. Iino, then modified by Dr. Usachev; EGFP-NFATc4, Addgene plasmid #10961).

    Techniques: Gene Expression, Transfection, Luciferase, Activity Assay, Concentration Assay

    Figure 2. NGF recruits TrkA but does not rely on the PLC pathway to activate NFAT-dependent gene expression. A. Schematic diagram of depictingtheexperimentalprotocols.B.AdiagramdescribingthePLCpathwaystimulatedbyNGF.C.Accordingtotheprotocoldescribed in (A), NGF (20 ng/ml, 5 h stimulation) was applied and its ability to activate NFAT-dependent gene expression in the presence or absence of K252a (a TrkA inhibitor) was assayed in PC12 cells as described in Figure 1. n = 6–7, ∗∗∗p < 0.001, One-Way ANOVA, Bonferroni post-hoc test D. U73122 (a PLC inhibitor) or Ca2+-dependent phosphatase calcineurin (CaN) inhibitors (FK506, CsA) were applied to block TrkA-triggered PLC activation or PLC-IP3-Ca2+ mediated CaN activation in PC12 cells treated as explained in (A). n = 6–7 E. NGF- treated PC12 cells were co-stimulated with KCl to depolarize cells for inducing an intracellular Ca2+ increase. To verify the integrity of CaN inhibitors, the inhibitors were applied to PC12 cells prepared as described in (A) to check if they inhibit CaN which should be activated by KCl. All inhibitors were applied 30 min prior to NGF stimulation. n = 6–7, ∗∗p < 0.01, ∗∗∗p < 0.001, One-Way ANOVA, Bonferroni post-hoc test. n stands for the number of well. Data are presented as mean ± S.E.M.

    Journal: All Life

    Article Title: NGF activates NFAT via the MEK1/2 pathway in PC12 cells

    doi: 10.1080/26895293.2022.2034670

    Figure Lengend Snippet: Figure 2. NGF recruits TrkA but does not rely on the PLC pathway to activate NFAT-dependent gene expression. A. Schematic diagram of depictingtheexperimentalprotocols.B.AdiagramdescribingthePLCpathwaystimulatedbyNGF.C.Accordingtotheprotocoldescribed in (A), NGF (20 ng/ml, 5 h stimulation) was applied and its ability to activate NFAT-dependent gene expression in the presence or absence of K252a (a TrkA inhibitor) was assayed in PC12 cells as described in Figure 1. n = 6–7, ∗∗∗p < 0.001, One-Way ANOVA, Bonferroni post-hoc test D. U73122 (a PLC inhibitor) or Ca2+-dependent phosphatase calcineurin (CaN) inhibitors (FK506, CsA) were applied to block TrkA-triggered PLC activation or PLC-IP3-Ca2+ mediated CaN activation in PC12 cells treated as explained in (A). n = 6–7 E. NGF- treated PC12 cells were co-stimulated with KCl to depolarize cells for inducing an intracellular Ca2+ increase. To verify the integrity of CaN inhibitors, the inhibitors were applied to PC12 cells prepared as described in (A) to check if they inhibit CaN which should be activated by KCl. All inhibitors were applied 30 min prior to NGF stimulation. n = 6–7, ∗∗p < 0.01, ∗∗∗p < 0.001, One-Way ANOVA, Bonferroni post-hoc test. n stands for the number of well. Data are presented as mean ± S.E.M.

    Article Snippet: SYBR green-based qPCR was performed with the cDNA from PC12 cells together with each NFAT plasmid DNA of known concentration (GFP-NFATc1, Addgene plasmid #11101; EGFPNFATc2, provided by Dr. Rao; EGFP-NFATc3, originally provided by Dr. Iino, then modified by Dr. Usachev; EGFP-NFATc4, Addgene plasmid #10961).

    Techniques: Gene Expression, Blocking Assay, Activation Assay

    Figure 3. NFAT activation by NGF depends on MEK1/2 but not PI3 K pathway. A. Schematic diagram of depicting the experimental pro- tocols. B. A diagram describing TrkA-mediated PI3 K and MEK1/2 activation by NGF. C-D. As described in Figure 2, NFAT reporter assays were performed with the exception that PI3 K inhibitor (LY294002, Wort (wortmannin) (C)) or MEK1/2 inhibitor (U0126, (D)) was applied to block TrkA-mediated PI3 K or MEK1/2 activation, respectively. All inhibitors were applied 30 min prior to NGF stimulation. n = 12–13 for (C), n = 6–12 for (D), ∗∗p < 0.01, ∗∗∗p < 0.001, One-Way ANOVA, Bonferroni post-hoc test. n stands for the number of well. Data are presented as mean ± S.E.M.

    Journal: All Life

    Article Title: NGF activates NFAT via the MEK1/2 pathway in PC12 cells

    doi: 10.1080/26895293.2022.2034670

    Figure Lengend Snippet: Figure 3. NFAT activation by NGF depends on MEK1/2 but not PI3 K pathway. A. Schematic diagram of depicting the experimental pro- tocols. B. A diagram describing TrkA-mediated PI3 K and MEK1/2 activation by NGF. C-D. As described in Figure 2, NFAT reporter assays were performed with the exception that PI3 K inhibitor (LY294002, Wort (wortmannin) (C)) or MEK1/2 inhibitor (U0126, (D)) was applied to block TrkA-mediated PI3 K or MEK1/2 activation, respectively. All inhibitors were applied 30 min prior to NGF stimulation. n = 12–13 for (C), n = 6–12 for (D), ∗∗p < 0.01, ∗∗∗p < 0.001, One-Way ANOVA, Bonferroni post-hoc test. n stands for the number of well. Data are presented as mean ± S.E.M.

    Article Snippet: SYBR green-based qPCR was performed with the cDNA from PC12 cells together with each NFAT plasmid DNA of known concentration (GFP-NFATc1, Addgene plasmid #11101; EGFPNFATc2, provided by Dr. Rao; EGFP-NFATc3, originally provided by Dr. Iino, then modified by Dr. Usachev; EGFP-NFATc4, Addgene plasmid #10961).

    Techniques: Activation Assay, Blocking Assay

    Figure 4. NFAc3 is the predominant NFAT isoform in PC12 cells. A. After constructing standard curves for each NFAT isoform, SYBR-based qPCR was performed with total RNA isolated from PC12 cells to calculate absolute copy number of each NFAT isoform. n = 3 B. Proposed model: NFATc3, a major NFAT isoform in PC12 cells shuttles in and out of the nucleus by CaN, an NFAT phosphatase and several kinases, respectively. An increase in NFAT-dependent gene expression by NGF depends on the MEK1/2 pathway via uncharacterized mechanisms in PC12 cells. Data are presented as mean ± S.E.M.

    Journal: All Life

    Article Title: NGF activates NFAT via the MEK1/2 pathway in PC12 cells

    doi: 10.1080/26895293.2022.2034670

    Figure Lengend Snippet: Figure 4. NFAc3 is the predominant NFAT isoform in PC12 cells. A. After constructing standard curves for each NFAT isoform, SYBR-based qPCR was performed with total RNA isolated from PC12 cells to calculate absolute copy number of each NFAT isoform. n = 3 B. Proposed model: NFATc3, a major NFAT isoform in PC12 cells shuttles in and out of the nucleus by CaN, an NFAT phosphatase and several kinases, respectively. An increase in NFAT-dependent gene expression by NGF depends on the MEK1/2 pathway via uncharacterized mechanisms in PC12 cells. Data are presented as mean ± S.E.M.

    Article Snippet: SYBR green-based qPCR was performed with the cDNA from PC12 cells together with each NFAT plasmid DNA of known concentration (GFP-NFATc1, Addgene plasmid #11101; EGFPNFATc2, provided by Dr. Rao; EGFP-NFATc3, originally provided by Dr. Iino, then modified by Dr. Usachev; EGFP-NFATc4, Addgene plasmid #10961).

    Techniques: Isolation, Gene Expression