Figure 7.

Klotho ameliorates calcification by promoting autophagy in MOVAS cells. (a) The use of KL significantly increases the LC3B-II/I ratio but decreases P62 protein expression in the CM + KL group compared with those in the CM group (n = 3). In addition, KL + CQ further promotes the LC3B-II/I ratio and restores P62 protein expression (n = 3) compared with those due to treatment with KL alone. (b) Both the number of yellow puncta and red puncta in merged images significantly increase in the CM group compared with those in the control group (n = 3). Moreover, the use of KL increases the number of free red and yellow puncta in merged images in the CM + KL group compared with those in the CM group (n = 3). The use of KL + CQ further increases the number of yellow puncta but decreases red puncta in merged images compared with those due to treatment with KL alone (n = 3). (c) In Alizarin Red S staining experiments, the use of KL ameliorates calcification staining in the CM + KL group compared with the CM group and the use of KL + CQ aggravates calcification staining compared with treatment with KL alone. (d) The Ca content significantly decreases after the use of KL compared with that of the CM group (n = 3) and markedly increases after the use of KL + CQ compared with that due to treatment with KL alone (n = 3). (e) The employment of KL significantly inhibits the Runx2 protein expression but promotes the α-SMA protein expression in the CM + KL group (n = 3) compared with those in the CM group, and the employment of KL + CQ significantly promotes the Runx2 protein expression and decreases the α-SMA protein expression (n = 3) compared with those due to treatment with KL alone. ^∗P < 0.05.