FIGURE SUMMARY
Title

Modulation of miR-204 Expression during Chondrogenesis

Authors
Dalle Carbonare, L., Bertacco, J., Minoia, A., Cominacini, M., Bhandary, L., Elia, R., Gambaro, G., Mottes, M., Valenti, M.T.
Source
Full text @ Int. J. Mol. Sci.
Figure 1

(A) MiR-204-5p and chondrogenic (SOX9, RUNX2, COMP, COL2A1) mRNA expression levels quantified by RT-PCR after 3, 7 and 14 days of chondrogenic differentiation in an MSCs in vitro model. RUNX2 protein expression levels assessed (B) by western blot analysis (left side) and quantified as a relative optical density ratio vs. B-actin (right side) on a protein lysate from an MSCs in vitro model after 3, 7 and 14 days of chondrogenic differentiation. (* p < 0.05; ** p < 0.005; *** p < 0.001 vs. expression levels of MSCs after 3 days of differentiation).
Figure 2

Protein levels assessed by western blot analysis (left side) and quantified as a relative optical density ratio vs. B-actin (right side) of (A) B-catenin and (B) ERK and pERK on protein lysates from MSCs after 3, 7 and 14 days of chondrogenic differentiation. (** p < 0.005; *** p < 0.001 vs. expression levels of MSCs after 3 days of differentiation).
Figure 3

miR-204-5p mRNA expression levels quantified by RT-PCR during (A) embryogenesis (3, 7 and 14 days post fecundation (dpf)) and (B) aging (2 and 6 months, 1 and 2 years old) in a zebrafish larvae in vivo model. (C) ERK, pERK and aggrecan protein levels assessed by western blot (top) and quantified as a relative optical density ratio vs. B-actin (down) on protein lysates from a zebrafish larvae in vivo model. (* p < 0.05; ** p < 0.005 vs. expression levels of larvae after 3 (dpf) (A,B) and vs. 1 year old (C)).
Figure 4

miR-204-5p, chondrogenic (SOX9, COL2A1, COMP, RUNX2) and osteogenic (RUNX2, COL1A1, IBSP, BGLAP, SPP1) mRNA expression levels quantified by RT-PCR during the early ((A,B) 3 days) and the middle ((C,D) 7 days) differentiation phases in miR-204-5p-silenced MSCs. (* p < 0.05; ** p < 0.005 vs. scramble/control).
Figure 5

miR-204-5p and osteogenic (RUNX2, COL1A1, IBSP, BGLAP, SPP1) mRNA expression levels quantified by RT-PCR at a late (10 days) differentiation phase in miR-204-5p-silenced MSCs. (* p < 0.05 vs. scramble/control).
Figure 6

miR-204-5p and SOX9 mRNA expression levels quantified by RT-PCR in (A) MSCs during chondrogenesis (7 days post differentiation) and in chondrocytes after interleukin 1 beta (IL 1 beta), (B) methylsulphonylmethane (MSM) and (C) clodronate treatment. (D) Cyclooxygenase 2 mRNA expression levels quantified by RT-PCR in MSCs during chondrogenesis (7 days post differentiation) and in chondrocytes with and without IL 1 beta and clodronate treatment. (* p < 0.05; ** p < 0.005 vs. controls).
Figure 7

Mechanism of action of miR-204-5p during the commitment and the differentiation phases of MSCs towards chondrogenesis or osteogenesis. miR-204-5p reduces MSCs’ commitment to the osteochondrogenic lineage and promotes the maturation of osteoblasts and chondroblasts by modulating the RUNX2 expression. The expression levels of RUNX2 during differentiation are shown in green.
Acknowledgments
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