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. 2021 Aug 17;9:681968. doi: 10.3389/fcell.2021.681968

FIGURE 6.

FIGURE 6

Overexpression of GPR65 stimulates activation of AMPK pathway. (A) Western blot analysis showed protein level of activated AMPK and activated JNK compared with their own total proteins in early OCPs cultured in DMEM or CT-26 CM, respectively. (B) Quantification of relative intensity of phosphorylation of AMPKα compared with total AMPKαin (A). (C) Quantification of relative intensity of phosphorylation of JNK compared with total JNK in (A). (D) Western blot analysis showed protein level of activated AMPK and activated JNK compared with their own total proteins in early OCPs after transfection with plasmid containing GPR65 with/without AMPKα siRNA in the presence of CT-26 CM. (E) Quantification of relative intensity of phosphorylation of AMPKα compared with total AMPKαin (D). (F) Quantification of relative intensity of phosphorylation of JNK compared with total JNK in (D). (G) Representative images of TRAP staining demonstrating osteoclast formation of early OCPs induced by RANKL and M-CSF after transfection with plasmids containing GPR65 alone or combination with AMPKα siRNA in the presence of CT-26 CM (scale bar = 250 μm). (H) Quantification of relative TRAP activity per well in (G). (I) Quantification of the number of osteoclasts per well in (G). ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.