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Kaplan-Meier survival curve comparing ( A ) male 129 vEDS <t>Map2k6</t> +/+ ( n = 17), vEDS Map2k6 +/– ( n = 20), and vEDS Map2k6 –/– mice ( n = 10) and ( B ) female 129 vEDS Map2k6 +/+ ( n = 19), vEDS Map2k6 +/– ( n = 15), and vEDS Map2k6 –/– mice ( n = 12). Significant differences were calculated using log-rank (Mantel-Cox) analysis (* P < 0.05; ** P < 0.01). ( C ) Immunoblot analysis of phosphorylated PKC at residue Ser 660 (pPKC) and phosphorylated ERK (pERK1/2) comparing aortic lysates obtained from the proximal descending aortas of mice at 2 months of age. ( D ) Quantification of p-PKC and p-ERK normalized to β-actin of control Map2k6 +/+ ( n = 3), vEDS Map2k6 +/+ ( n = 4), control Map2k6 –/– ( n = 7), and vEDS Map2k6 –/– ( n = 5) mice. P value refers to 2-way ANOVA with Holm-Šídák post hoc test (* P < 0.05, *** P < 0.001). ( E ) Immunofluorescence of sections from the proximal descending thoracic aorta of vEDS Map2k6 +/+ and vEDS Map2k6 –/– mice. The dashed line marks the approximate boundaries of the aortic wall. Scale bar is 50 microns. ( F ) Mean and total protein phosphatase 1 (PP1) dephosphorylation activity in aortic protein lysates from 129 vEDS Map2k6 +/+ mice ( n = 6) and 129 vEDS Map2k6 –/– ( n = 7). P value refers to unpaired t test with Welch’s correction (* P < 0.05, ** P < 0.01).DiFMU, 6,8-difluoro-7-hydroxy-4-methylcoumarin. In D and F , each symbol represents an independent biological replicate, with unfilled symbols representing male samples. Error bars show mean ± SEM. ( G ) Kaplan-Meier survival curve comparing control 129 vEDS Map2k6 –/– mice ( n = 33, 17 females and 16 males) with 129 vEDS Map2k6 –/– ( n = 11, 8 females and 3 males) mice receiving ruboxistaurin (PKC inhibitor) starting at postnatal day 21. Significant differences were calculated using log-rank (Mantel-Cox) analysis (* P < 0.05).
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Kaplan-Meier survival curve comparing ( A ) male 129 vEDS Map2k6 +/+ ( n = 17), vEDS Map2k6 +/– ( n = 20), and vEDS Map2k6 –/– mice ( n = 10) and ( B ) female 129 vEDS Map2k6 +/+ ( n = 19), vEDS Map2k6 +/– ( n = 15), and vEDS Map2k6 –/– mice ( n = 12). Significant differences were calculated using log-rank (Mantel-Cox) analysis (* P < 0.05; ** P < 0.01). ( C ) Immunoblot analysis of phosphorylated PKC at residue Ser 660 (pPKC) and phosphorylated ERK (pERK1/2) comparing aortic lysates obtained from the proximal descending aortas of mice at 2 months of age. ( D ) Quantification of p-PKC and p-ERK normalized to β-actin of control Map2k6 +/+ ( n = 3), vEDS Map2k6 +/+ ( n = 4), control Map2k6 –/– ( n = 7), and vEDS Map2k6 –/– ( n = 5) mice. P value refers to 2-way ANOVA with Holm-Šídák post hoc test (* P < 0.05, *** P < 0.001). ( E ) Immunofluorescence of sections from the proximal descending thoracic aorta of vEDS Map2k6 +/+ and vEDS Map2k6 –/– mice. The dashed line marks the approximate boundaries of the aortic wall. Scale bar is 50 microns. ( F ) Mean and total protein phosphatase 1 (PP1) dephosphorylation activity in aortic protein lysates from 129 vEDS Map2k6 +/+ mice ( n = 6) and 129 vEDS Map2k6 –/– ( n = 7). P value refers to unpaired t test with Welch’s correction (* P < 0.05, ** P < 0.01).DiFMU, 6,8-difluoro-7-hydroxy-4-methylcoumarin. In D and F , each symbol represents an independent biological replicate, with unfilled symbols representing male samples. Error bars show mean ± SEM. ( G ) Kaplan-Meier survival curve comparing control 129 vEDS Map2k6 –/– mice ( n = 33, 17 females and 16 males) with 129 vEDS Map2k6 –/– ( n = 11, 8 females and 3 males) mice receiving ruboxistaurin (PKC inhibitor) starting at postnatal day 21. Significant differences were calculated using log-rank (Mantel-Cox) analysis (* P < 0.05).

Journal: JCI Insight

Article Title: Map2k6 is a potent genetic modifier of arterial rupture in vascular Ehlers-Danlos syndrome mice

doi: 10.1172/jci.insight.187315

Figure Lengend Snippet: Kaplan-Meier survival curve comparing ( A ) male 129 vEDS Map2k6 +/+ ( n = 17), vEDS Map2k6 +/– ( n = 20), and vEDS Map2k6 –/– mice ( n = 10) and ( B ) female 129 vEDS Map2k6 +/+ ( n = 19), vEDS Map2k6 +/– ( n = 15), and vEDS Map2k6 –/– mice ( n = 12). Significant differences were calculated using log-rank (Mantel-Cox) analysis (* P < 0.05; ** P < 0.01). ( C ) Immunoblot analysis of phosphorylated PKC at residue Ser 660 (pPKC) and phosphorylated ERK (pERK1/2) comparing aortic lysates obtained from the proximal descending aortas of mice at 2 months of age. ( D ) Quantification of p-PKC and p-ERK normalized to β-actin of control Map2k6 +/+ ( n = 3), vEDS Map2k6 +/+ ( n = 4), control Map2k6 –/– ( n = 7), and vEDS Map2k6 –/– ( n = 5) mice. P value refers to 2-way ANOVA with Holm-Šídák post hoc test (* P < 0.05, *** P < 0.001). ( E ) Immunofluorescence of sections from the proximal descending thoracic aorta of vEDS Map2k6 +/+ and vEDS Map2k6 –/– mice. The dashed line marks the approximate boundaries of the aortic wall. Scale bar is 50 microns. ( F ) Mean and total protein phosphatase 1 (PP1) dephosphorylation activity in aortic protein lysates from 129 vEDS Map2k6 +/+ mice ( n = 6) and 129 vEDS Map2k6 –/– ( n = 7). P value refers to unpaired t test with Welch’s correction (* P < 0.05, ** P < 0.01).DiFMU, 6,8-difluoro-7-hydroxy-4-methylcoumarin. In D and F , each symbol represents an independent biological replicate, with unfilled symbols representing male samples. Error bars show mean ± SEM. ( G ) Kaplan-Meier survival curve comparing control 129 vEDS Map2k6 –/– mice ( n = 33, 17 females and 16 males) with 129 vEDS Map2k6 –/– ( n = 11, 8 females and 3 males) mice receiving ruboxistaurin (PKC inhibitor) starting at postnatal day 21. Significant differences were calculated using log-rank (Mantel-Cox) analysis (* P < 0.05).

Article Snippet: Map2k6 –/– mice were genotyped according to The Jackson Laboratory protocols.

Techniques: Western Blot, Residue, Control, Immunofluorescence, De-Phosphorylation Assay, Activity Assay