AT1aR deficiency did not cause alterations of AT1bR and AT2R in adipose tissue.

A. Gene expression of AT1bR in adipose tissue. B. Protein expression of AT2R in adipose tissue. Data were presented as Mean ± S.E.M. n = 4. (TIF)</p

AT1aR knockout improved epididymal histomorphology induced by high fat diet.

A, B: Body weight of rats was recorded weekly and body weight gain was calculated by weight increase in 12 weeks. C. Daily food intake of rats. D: Physical morphology of epididymal fat. E: Epididymal fat index (showed as epididymal fat weight/body weight). F: Adipocytes morphology was visualized by HE staining and adipocyte area were calculated by Image J. *p +p -/—ND rats; Ψp < 0.05 vs WT-ND rats. Data were presented as Mean ± S.E.M. n = 6.</p

sgRNA primer sequences for AT1aR^-/- rats generation.

sgRNA primer sequences for AT1aR-/- rats generation.</p

Primer sequences used for quantitative real time RT-PCR.

Primer sequences used for quantitative real time RT-PCR.</p

Development of AT1aR^-/- rats.

A. sgRNA combined CRISPR/Cas system to generate AT1aR-/- rats. B, C. PCR results for AT1aR-/- rats and WT rats. M: marker; N: negative control; P: positive control D. AT1aR deficiency was confirmed in major Ang II responsive tissues. Data were presented as Mean ± S.E.M. n = 5. (TIF)</p

AT1aR knockout improved insulin resistance and metabolic disorders in high-fat diet rats.

A: Oral glucose tolerance test (OGTT) curve and area under the curve. B: Insulin tolerance test (ITT) curve and area under the curve. C: Fasting blood glucose (FBG). D: Blood pressure. E: Serum triglyceride (TG) levels. F: Serum total cholesterol (T-CHO). G: Serum free fatty acids (NEFAs). H: Serum glycerol. I: Serum low-density lipoprotein (LDL). J: Serum high-density lipoprotein (HDL). *p #p < 0.05 vs WT-HFD rats. Data were presented as Mean ± S.E.M. n = 6.</p

AT1aR knockout activated cAMP/PKA pathway.

A: Protein expressions of PKA, P-GSK-3β (ser9) and GSK-3β in adipose tissue. B: cAMP levels in adipose tissue. C: The proposed pathway for Ang II inhibiting adipose lipolysis. By binding to AT1R, Ang II activates inhibitory Gi which in turn reduces cAMP production. As a result, PKA activation is limited and HSL phosphorylation is decreased, leading to inhibitory of triglyceride hydrolysis. +p -/—ND rats. Data were presented as Mean ± S.E.M. n = 6.</p

High fat diet activated Ang II-AT1R axis in rats.

A. Protein expressions of kinases downstream of the Ang II-AT1R axis, p38 and ERK. B. Gene expression of Nox4 in adipose tissue. *p < 0.05 vs WT-ND rats. Data were presented as Mean ± S.E.M. n = 4.</p

AT1aR knockout promoted adipose lipolysis.

A, B: Gene expressions of key enzymes for lipid synthesis in adipose tissue, FAS (A) and ACC (B). C: Protein expressions of adipose lipolysis related enzymes, ATGL and HSL. D, E: Levels of NEFAs (D) and glycerol (E) in culture medium released by adipose tissue within 24 hours. *p +p -/—ND rats. Data were presented as Mean ± S.E.M. n = 6.</p

S1 Raw images -

(PDF)</p