metabolic reprogramming in PMOP

1.Morphological data
Micro-CT and morphological data in SHAM, OVX and OVX+ICA group as well as in OVX, OVX+low-dosage DCA and OVX+low-dosage DCA group.

2. The Rat Glucose Metabolism RT² Profiler PCR Array
84 glucose metabolism related genes were detected in SHAM, OVX and OVX+ICA groups.

3. Immunoblotting raw pictures
For detections in SHAM, OVX and OVX+ICA groups, the sequences in the band is SHAM, OVX and ICA+OVX from left to right.For detections in cells, the sequences in the band is siCON, siPDK1, DCA and ICA from left to right. For detections in DCA-interventioned OVX group, the sequences in the band is OVX, low-dosage DCA and high-dosage DCA from left to right. For PDK1 enzymatic activity detection, the sequences in the band was phosphorylated PDHA Ser232 at 0, 5, 10 minutes.

4. Immunofluorescence pictures
Immunofluorescences of PDHA and PDHA pSer232 were performed on paraffin cuts in SHAM, OVX and OVX+ICA group, reconfirming the upregulation of PDHA pSer232 in OVX group.
The co-labelled PDHA pSer232 with osteoclast marker and osteoblast marker respectively indicated the the upregulation of PDHA pSer232 mainly existed in osteoclasts.

5.qPCR raw data
Osteoclast marker genes (Nfatc1, Tracp, Oscar, Mmp9 and Cathepsin K ) significantly decreased after PDK1 knockdown, DCA and ICA administration, indicating that DCA and ICA both hindered osteoclast differentiation.

6.TRAP staining pictures
The numbers of TRAP positive osteoclasts dramatically diminished after PDK1 knockdown, DCA and ICA administration.