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. 2021 Feb 3;128(6):755–768. doi: 10.1161/CIRCRESAHA.120.316666

Figure 6.

Figure 6.

Calcineurin (PP2B) regulates VASPS239 phosphorylation after vascular smooth muscle (VSM) bcl11b (B-cell leukemia 11b) deletion. A, Representative Western blot images demonstrating increased PP2B expression, increased NFAT2 and impaired VASP (vasodilator-stimulated phosphoprotein) phosphorylation at serine 239 (pVASPS239) in tamoxifen-inducible VSM-specific Bcl11b null mice (BSMKO) VSM cells compared with wild-type (WT) controls. GAPDH serves as loading control. Each lane represents a cell preparation from one mouse for a total of 4 replicates. B, Treatment of BSMKO VSM cells with 1 or 10 μmol/L cyclosporine A (CsA), a calcineurin inhibitor, or (C) a specific calcineurin autoinhibitory peptide (CAIP; 10 or 100 μmol/L) reversed pVASPS239 in BSMO cells towards WT levels, in a dose-dependent manner. V, vehicle control. Quantitation of band intensities in graph (n=3 replicate experiments). *P=1.5×10−2 vs WT/vehicle; †P=3.0×10−2 vs BSMKO/vehicle by 1-way ANOVA with Tukey multiple comparisons test. D, Western blots for Bcl11b, PKG1 (PKG, isoform 1), PP2B and pVASPS239 in WT and BSMKO aortic media (adventitia removed) without (left) or with (right) transient transfection with vehicle (lipofectamine C, control) or 20 μg Bcl11b plasmid. GAPDH serves as loading control.