Figure 2. Adp Inhibits Murine Adipogenesis.

(A) qPCR analysis of Adp expression levels in various tissues from 4-month old ICR mice. Brown adipose tissue (BAT), inguinal white adipose tissue (IWAT), perigonadal white adipose tissue (GWAT), mesenteric white adipose tissue (MWAT)

(B) 3T3-L1 cells were induced to form adipocytes and Adp levels were assessed with qPCR.

(C) Adp expression was quantified with qPCR in stromal-vascular (SV) and adipocyte (Adipo) fractions.

(D) Adp expression levels in adipose depots of littermates provided a normal or high fat diet (HFD) (left panel), genetically obese (ob/ob) fat depots compared to littermate controls (middle panel), or mice subjected to a 24-hour fast compared to fed controls (right panel).

(E) Adp levels in 3T3-L1 cells infected with virus encoding GFP or Adp.

(F–H) 3T3-L1 cells were infected with virus encoding GFP or Adp, adipogenically induced, and adipogenesis was evaluated with (F) Oil Red O staining (fat stains red), (G) triglyceride quantitation and (H) qPCR quantitation of the indicated markers. PPARγ and C/EBPα are adipogenic transcription factors; aP2 marks differentiated adipocytes; adipsin and leptin are adipokines expressed by mature adipocytes; Pref-1 is a preadipocyte marker.

(I) NIH3T3 cells were infected with virus encoding GFP or Adp, adipogenically induced, and adipogenesis evaluated with Oil Red O staining.

(J, K) MC3T3-E1 preosteoblastic cells were infected with GFP or Adp, osteogenically induced, and bone formation was assessed with (J) Von Kossa staining (bone stains black) or (K) qPCR of osteogenic markers. Runx2 (Runx) and Osterix (Osx) are osteogenic transcription factors, and alkaline phosphatase (Alp) and osteocalcin (OC) are bone differentiation markers.

*p < 0.05, **p < 0.01; N/S not significant by t-test. Error bars represent SEM. β-actin serves as a loading control.