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. Author manuscript; available in PMC: 2022 Oct 29.
Published in final edited form as: Circ Res. 2021 Sep 23;129(10):e202–e214. doi: 10.1161/CIRCRESAHA.121.319374

Figure 6: ADAR1 regulated SMC phenotype and MMP level/activity independent of its editing function.

Figure 6:

A-I, Mouse aortic SMCs were transduced with control (−) or ADAR1 shRNA (shADAR1) adenoviral vector and then treated with 10 ng/ml of IL-1β for 24 h. Protein expressions of ADAR1, inflammatory cell marker VCAM-1, MMP2, MMP9, and SMC markers were detected by Western blot and quantified by normalizing to GAPDH, respectively. MMP activity was measured by in situ zymography (I). *P =1.45E-02 for ADAR1 110 kD, 2.05E-02 for ADAR1 150 kD (B), 2.08E-03 (C), 2.85E-03 for pre-MMP2, 1.45E-03 for m-MMP2 (D), 1.85E-03 for pre-MMP9, 2.35E-03 for m-MMP9 (E), 2.11E-02 (F), 1.41E-02 (G), 8.41E-03 (H), 1.85E-03 (I) vs. vehicle-treated cells (−), respectively; #P = 1.98E-02 for ADAR1 110 kD, 2.48E-02 for ADAR1 150 kD (B), 3.11E-02 (C), 2.53E-02 for pre-MMP2, 1.83E-02 for m-MMP2 (D), 2.53E-02 for pre-MMP9, 3.23E-02 for m-MMP9 (E), 4.05E-02 (F), 2.05E-02 (G), 1.35E-02 (H), 2.53E-02 (I) vs. control adenovirus-transduced cells (−) with IL-1β treatment, respectively. n=6 for B-H; n=3 for I. J-L, Mouse aortic SMCs were transduced with control (−) or ADAR1 shRNA adenoviral vector (shADAR1) and then treated with vehicle (−) or 10 ng/ml of IL-1β for 24 h. The precursor (pre-) and mature (m)-mRNA levels of MMP2 and MMP9 were detected by RT-PCR (J). The pre- and m-mRNA levels of MMP2 (K) and MMP9 (L) were quantified by normalizing to GAPDH, respectively. *P = 4.92E-03 for pre-MMP2, 1.99E-02 for m-MMP2 (K) and 4.92E-03 for pre-MMP9, 9.22E-03 for m-MMP9 (L) vs. vehicle-treated cells (−); #P = 5.98E-03 for pre-MMP2, 3.40E-02 for m-MMP2 (K) and 2.98E-02 for pre-MMP9, 3.18E-02 for m-MMP9 (L) vs. control vector-transduced cells (−) with IL-1β treatment; n=6. All protein or mRNA levels were shown as fold induction relative to the mean value of vehicle (−) and control vector-transduced cells (−), which was set as 1 for each comparison. Kruskal-Wallis test with Dunn’s multiple comparisons tests were performed to determine statistical difference for all panels.