Fig 1: Knockdown of IPMK restores the induction of autophagy and activation of the AMPK/ULK1 signaling pathway by ebastine treatment. (a) Expression of LC3B and p-AMPK was analyzed by western blot after knockdown of 10 genes in MNNG cells. (b) After knockdown of IPMK in MNNG cells, the mRNA expression of IPMK was determined by qRT-PCR. (c) Protein expression of IPMK and AMPK was analyzed by western blot after knockdown of IPMK in MNNG cells. (d-f) Osteosarcoma cells were treated with ebastine for 48 h with or without knockdown of IPMK, and LC3 puncta were analyzed by the mRFP-GFP-LC3 construct. (g) Protein levels of apoptosis, autophagy, IPMK, AMPK and p-AMPKK in osteosarcoma cells treated with ebastine for 48 h with or without knockdown of IPMK. Data are shown as the means ± SD from at least three independent experiments. Statistical analysis was performed using Student's t-test. Error bars represent the SEM. *P<0.05.
Fig 2: Ebastine exerts antitumor activity through the IPMK/AMPK/ULK1 signaling pathway in vivo. (a) Protein expression of IPMK, AMPK, p-AMPK, ULK1, LC3B, and CDK2 and cleavage of caspase-9 were analyzed by western blot in tumor tissues. (b-d) IHC staining of related proteins. (e) The diagram of the mechanism of ebastine anti-osteosarcoma. Statistical analysis was performed using Student's t-test. Error bars represent the SEM. *P<0.05, ** P<0.01; *** P<0.001.
Supplier Page from Abcam for Anti-IPMK antibody