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. 2019 Aug 28;16(6):1092–1110. doi: 10.1080/15548627.2019.1659612

Figure 7.

Figure 7.

Reduced SQSTM1 expression increases ROS levels and senescence of VSMCs. RASMs were transfected with siRNA control or siSqstm1 for measurements of SQSTM1 and CDKN1A expression (A-C), and SA-GLB1 activity, using FDG, in response to 100 nM AGT II or 50 μM zinc for 3 d (D). ppargc1a−/- VSMCs were infected with 1 × 107 PFU of adenovirus control (Ad.Control) or adenovirus containing human SQSTM1 gene (Ad.SQSTM1) for ROS measurements (E) and SA-GLB1 activity using FDG (F). Ppargc1a+/+ and ppargc1a−/- VSMCs infected with Ad.Control or Ad.SQSTM1 were lysed for analysis of SQSTM1 and LC3-II expression. (G-I). Expression was expressed as fold change compared with siControl-transfected cells. Aortas of 2-month-old mice Sqstm1+/+, Sqstm1± and sqstm1−/- in the C57Bl/6 background (8 males each) were cleaned, fixed and tested for SA-GLB1 activity using FDG (J). Fluorescence was adjusted by the weight of aortas. * denotes p < 0.01. Body weight for each genotype is expressed in grams (g) (K). Aortas of each genotype (2-month-old, males) were cleaned of periadventitial fat, lysed and 80 μg of total extracts were separated in a 4–20% precast Criterion gels (L). Quantification of TRP53 and LC3-II adjusted by ACTB is shown in M and N, respectively. * denotes p < 0.05.