Supplementary Figure Legends
Supplemental Figure S1. Trabecular bone loss and bone marrow fat increase in osteoporosis
femur. (A-D): Trabecular bone loss in osteoporosis femur. The femurs from Sham and OVX mice
were scanned by Micro-CT three month after surgery. Representative pictures of the coronal and
horizontal plane of femur were shown (A). The BMD (B) and morphologic factor of trabecular
bone (C, D) were calculated. (E): The weight of Sham and OVX mice were calculated. (F, G):
Bone marrow fat increased in osteoporosis femur. The bone marrow fat was detected by toluidine
blue staining (F) and the adipocyte size was quantified with Image Pro software (G). Data are
shown as means ± SD. *P < 0.05, vs. Sham, n = 3. Scale bar in micrographs represents 200μm.
Abbreviations: Sham, sham surgery; OVX, ovariectomy; BMD, bone mineral density; Tb.N,
trabecular bone number; Tb.Th, trabecular bone thickness.
Supplemental Figure S2. Cell surface markers expression in MSCs from Sham and OVX bone
marrow. Flow-cytometry was performed to determine the surface markers expressed in Sham
MSCs (A) and OVX MSCs (B).
Supplemental Figure S3. Effect of mimics and inhibitor to regulate miR-705 and miR-3077-5p
expression. (A) Negative control, miR-705 mimics and inhibitor were transfected in MSCs for 48
hours. Real-time RT-PCR was performed to determine the expression of miR-705. (B) Negative
control, miR-3077-5p mimics and inhibitor were transfected in MSCs for 48 hours. Real-time
RT-PCR was performed to determine the expression of miR-3077-5p. Data are shown as means ±
SD. *P < 0.05, **P<0.01, n = 3.
Supplemental Figure S4. Effect of mimics and inhibitor to regulate miR-705 and miR-3077-5p
expression during induction. (A) The transfected MSCs was cultured in osteogenic medium for 14
days. Real-time RT-PCR was performed to detect the expression of miR-705 and miR-3077-5p
after osteogenic induction. (B) The transfected MSCs was cultured in adipogenic medium for 7
days. Real-time RT-PCR was performed to detect the expression of miR-705 and miR-3077-5p
after adipogenic induction. Data are shown as means ± SD. *P < 0.05, **P<0.01, n = 3.
Supplemental Figure S5. Effect of IKKα SiRNA to regulate IKKα expression. (A, B): Negative
control and IKKα SiRNA were transfected in MSCs for 48 hours. Real-time RT-PCR was
performed to determine the expression of IKKα (A). Western blot was performed to determine the
expression of P65 (B). Data are shown as means ± SD. *P < 0.05, **P<0.01, n = 3.
Supplemental table
Supplemental table S1. Primer sequence used for real-time RT-PCR detection
Gene
Primer Sequence
Forward
Reverse
β-actin
5'-CTGGCACCACACCTTCTACA-3'
5'-GGTACGACCAGAGGCATACA-3'
LPL
5'-CCCCAGTCGCCTTTCTCCTGAT-3'
5'-CTCTTGGCTCTGACCTTGTTGAT-3'
PPARγ
5'-ACTGCCGGATCCACAAAA-3'
5'-TCTCCTTCTCGGCCTGTG-3'
OCN
5'-CTGACAAAGCCTTCATGTCCAA-3'
5'-GCGCCGGAGTCTGTTCACTA-3'
RUNX2
5'-GACTGTGGTTACCGTCATGGC-3'
5'-ACTTGGTTTTTCATAACAGCGGA-3'
HOXA10
5'-TTCGCCGGAGAAGGACTC-3'
5'-TCTTTGCTGTGAGCCAGTTG-3'
IKKα
5'-GTGAACATCCTCTGACATGTGTGGT-3'
5'-GCAACACAAGGAGGCTGGGCT-3'