Addgene inc pcaf expression plasmid

Figure 4 | ERK retrograde signalling controls <t>PCAF</t> activation. (a,b) NGF stimulates pERK, PCAF and H3K9ac expressions in adult DRG cultures after 3-h treatment, which is abrogated by the ERK kinase inhibitor PD98059 (PD), ICC (a) and fold change analysis of intensity density (b). Scale bar, 20 mm, N ¼ 3 per group, performed in triplicate. (c,d) Nuclear PCAF immunoprecipitation from in vivo L4-L6 DRG 24 h following Sham or SNA reveals an increase in <t>PCAF</t> <t>expression</t> and threonine phosphorylation following SNA but not serine phosphorylation, immunoblot (c) and fold change of density analysis (d). N ¼ 5 per group, performed in triplicate. (e–i) In L4-L6 DRG, 24 h following SNA we observe an increase in pERK (e,f), PCAF (g,h) and H3K9ac (g,i) expression, which is signiﬁcantly decreased by ERK inhibition with PD at the nerve stump. Insert shows high nuclear expression of PCAF and H3K9ac after SNA. Scale bars, 75 mm, N ¼ 3 per group, performed in triplicate. (j–l) PD also inhibits gene expression (Q-PCR, N ¼ 3 per group) (j) as well as H3K9ac (k) and PCAF (l) at the promoters of GAP-43, Galanin and BDNF 24 h following SNA (ChIPs). N ¼ 6 per group, performed in triplicate. Error bars, s.e. (b,f,h,i) Po0.0001, ANOVA, Bonferroni post hoc tests, **Po0.001 and ***Po0.001, (d,j–l) Student’s t-test, *Po0.05, **Po0.001 and ***Po0.001. Original immunoblot images are shown in Supplementary Fig. 12.

Pcaf Expression Plasmid, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Figure 4 | ERK retrograde signalling controls PCAF activation. (a,b) NGF stimulates pERK, PCAF and H3K9ac expressions in adult DRG cultures after 3-h treatment, which is abrogated by the ERK kinase inhibitor PD98059 (PD), ICC (a) and fold change analysis of intensity density (b). Scale bar, 20 mm, N ¼ 3 per group, performed in triplicate. (c,d) Nuclear PCAF immunoprecipitation from in vivo L4-L6 DRG 24 h following Sham or SNA reveals an increase in PCAF expression and threonine phosphorylation following SNA but not serine phosphorylation, immunoblot (c) and fold change of density analysis (d). N ¼ 5 per group, performed in triplicate. (e–i) In L4-L6 DRG, 24 h following SNA we observe an increase in pERK (e,f), PCAF (g,h) and H3K9ac (g,i) expression, which is signiﬁcantly decreased by ERK inhibition with PD at the nerve stump. Insert shows high nuclear expression of PCAF and H3K9ac after SNA. Scale bars, 75 mm, N ¼ 3 per group, performed in triplicate. (j–l) PD also inhibits gene expression (Q-PCR, N ¼ 3 per group) (j) as well as H3K9ac (k) and PCAF (l) at the promoters of GAP-43, Galanin and BDNF 24 h following SNA (ChIPs). N ¼ 6 per group, performed in triplicate. Error bars, s.e. (b,f,h,i) Po0.0001, ANOVA, Bonferroni post hoc tests, **Po0.001 and ***Po0.001, (d,j–l) Student’s t-test, *Po0.05, **Po0.001 and ***Po0.001. Original immunoblot images are shown in Supplementary Fig. 12.

Journal: Nature communications

Article Title: PCAF-dependent epigenetic changes promote axonal regeneration in the central nervous system.

doi: 10.1038/ncomms4527

Figure Lengend Snippet: Figure 4 | ERK retrograde signalling controls PCAF activation. (a,b) NGF stimulates pERK, PCAF and H3K9ac expressions in adult DRG cultures after 3-h treatment, which is abrogated by the ERK kinase inhibitor PD98059 (PD), ICC (a) and fold change analysis of intensity density (b). Scale bar, 20 mm, N ¼ 3 per group, performed in triplicate. (c,d) Nuclear PCAF immunoprecipitation from in vivo L4-L6 DRG 24 h following Sham or SNA reveals an increase in PCAF expression and threonine phosphorylation following SNA but not serine phosphorylation, immunoblot (c) and fold change of density analysis (d). N ¼ 5 per group, performed in triplicate. (e–i) In L4-L6 DRG, 24 h following SNA we observe an increase in pERK (e,f), PCAF (g,h) and H3K9ac (g,i) expression, which is signiﬁcantly decreased by ERK inhibition with PD at the nerve stump. Insert shows high nuclear expression of PCAF and H3K9ac after SNA. Scale bars, 75 mm, N ¼ 3 per group, performed in triplicate. (j–l) PD also inhibits gene expression (Q-PCR, N ¼ 3 per group) (j) as well as H3K9ac (k) and PCAF (l) at the promoters of GAP-43, Galanin and BDNF 24 h following SNA (ChIPs). N ¼ 6 per group, performed in triplicate. Error bars, s.e. (b,f,h,i) Po0.0001, ANOVA, Bonferroni post hoc tests, **Po0.001 and ***Po0.001, (d,j–l) Student’s t-test, *Po0.05, **Po0.001 and ***Po0.001. Original immunoblot images are shown in Supplementary Fig. 12.

Article Snippet: PCAF expression plasmid was obtained from Addgene (Plasmid 8941).

Techniques: Activation Assay, Immunoprecipitation, In Vivo, Expressing, Phospho-proteomics, Western Blot, Inhibition, Gene Expression

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