Journal: JCI Insight

Article Title: Opposing roles for myeloid and smooth muscle cell STING in pulmonary hypertension

doi: 10.1172/jci.insight.184792

Figure Lengend Snippet: ( A ) RVSP measurements of littermate control (WT) and endothelial cell–specific STING-deficient (eSTING) mice exposed to normoxia or chronic hypoxia. ( B ) Fulton index of WT and eSTING mice from the indicated experimental groups. ( C ) RVSP curve representation and ( D ) RVSP measurement of littermate control (WT) and smooth muscle–specific STING-deficient (smSTING-deficient) mice exposed to normoxia or chronic hypoxia. ( E ) Fulton index of WT and smSTING mice in the indicated experimental groups. ( F ) Representative images of α-SMA (brown, arrowheads) IHC staining, with insets, of formalin-fixed lung sections from mice across indicated experimental groups. Scale bars: 100 μm. Original magnification ×10. ( G and H ) Quantification of ( G ) small, medium, and large and ( H ) completely or partially muscularized pulmonary vessels of WT and smSTING mice across experimental groups, as assessed through α-SMA IHC staining. ( I and J ) Quantification of Western blot of ( I ) TIMP3 and ( J ) MMP8 of smSTING and littermate control mice from different experimental groups. Each dot represents an individual mouse ( n = 4–10/group). ( K ) Corresponding Western blots from mice in the indicated experimental groups ( n = 4/group). ( L ) Gelatin zymography detection of pro-MMP2, MMP2, pro-MMP9, and MMP9 in lysates from isolated vascular smooth muscle cells (Vascular SMC) exposed to hypoxic conditions (10% ambient O 2 ) for 24 hours ( n = 4/group). Data are presented as mean ± SEM. Significance levels were calculated with ANOVA followed by unpaired 2-tailed Welch’s t test corrected for multiple comparisons by use of Dunnett’s test. P values are shown in the graphs. P values in G were calculated from total vessels (small + medium + large) of mice from the experimental groups, unless specified otherwise.

Article Snippet: Membranes were blocked with EveryBlot Blocking Buffer (Bio-Rad 12010020) for 10 minutes at room temperature, then incubated with primary antibodies (1:1,000) against MMP8 (ProteinTech 17874-1), MMP9 (Cell Signaling Technology 24317AS), MMP2 (Cell Signaling Technology 87809S), TIMP3 (Cell Signaling Technology 5673S), STING (Cell Signaling Technology 13647), and β-Actin (Cell Signaling Technology 4967L) at 4°C overnight.

Techniques: Control, Immunohistochemistry, Western Blot, Zymography, Isolation