Fig 1: Exosomal HOTTIP contributed to cisplatin resistance by upregulating HMGA1 in GC cells. (A and B) The protein level of HMGA1 was evaluated in MGC-803 and MKN-45 cells were incubated with PBS, exosomes, exosomes + si-NC, or exosomes + si-HOTTIP#1 using Western blot analysis. (C-J) MGC-803 and MKN-45 cells were transfected with si-NC, si-HMGA1, si-HMGA1 + pcDNA, si-HMGA1 +HOTTIP. (C and D) Western blot was performed to measure the protein level of HMGA1. (E and F) CCK-8 assay was utilized to assess cell proliferation capacity. (G and H) Transwell was employed to examine migration and invasion capacities in transfected MGC-803 and MKN-45 cells. (I and J) The protein abundances (E-cadherin, N-cadherin and Vimentin) were evaluated using the Western blot assay. *P<0.05.
Fig 2: HOTTIP regulated HMGA1 through functioning as a ceRNA of miR-218 in GC cells. (A and B) The qRT-PCR analysis was performed to evaluate the subcellular location of HOTTIP in GC cells. (C) Potential binding sites of miR-218 within HOTTIP or HMGA1 were predicted using the miRcode or starBase, respectively. (D) The qRT-PCR assay was carried out to analyze the abundance of miR-218 in cisplatin-resistant and -sensitive GC cells. (E and F) Dual-luciferase reporter assay was conducted to determine the luciferase activity in MGC-803 and MKN-45 cells co-transfected with miR-NC or miR-218 and WT-HOTTIP or MUT-HOTTIP. (G and H) The luciferase activity was measured in MGC-803 and MKN-45 cells co-transfected with miR-NC or miR-218 and HMGA1 3ʹUTR-WT or HMGA1 3ʹUTR-MUT. (I and J) The enrichment of HOTTIP, miR-218, and HMGA1 was measured by RIP assay in MGC-803 and MKN-45 cells incubated with Ago2 or IgG. (K and L) The protein level of HMGA1 was detected in MGC-803 and MKN-45 cells transfected with miR-NC, miR-218, miR-218+ pcDNA, or miR-218+ HOTTIP using the Western blot analysis. *P<0.05.
Supplier Page from Abcam for Anti-HMGA1 antibody