Fig 1: Klotho mitigates H/R injury by regulating the Hsp70/Akt/Bad pathway. (A) H9c2(2-1) cells were transfected with klotho plasmid for 24 h prior to H/R injury. Expression levels of klotho and Hsp70 were detected by immunofluorescence microscopy (magnification, x200). DAPI was used to stain the nuclei. (B) Expression of klotho, p-Akt, p-Bad, total Akt and total Bad was detected by western blotting. ß-actin was used as the internal reference gene. (C) H9c2(2-1) cells were infected with klotho shRNA lentivirus or control shRNA lentivirus for 48 h. The levels of klotho, p-Akt and total Akt were detected by western blotting. H/R, hypoxia/reoxygenation; Ctrl, control; Hsp70, heat shock protein 70; p-, phosphorylated; sh, short hairpin.