Journal: Scientific Reports
Article Title: Mitragynine, an euphoric compound inhibits hERG1a/1b channel current and upregulates the complexation of hERG1a-Hsp90 in HEK293-hERG1a/1b cells
Figure Lengend Snippet: The effects of ( A,B ) arsenic trioxide and ( C,D ) mitragynine on the hERG1a/1b protein expression. Representative image of Western blot analysis for the protein expression of hERG1a (155 kDa and 135 kDa), hERG1b (95 kDa and 80 kDa) and β-actin (43 kDa) following treatment with arsenic trioxide or mitragynine for 24 h. Electrophoresis was carried out at 150 V for 90 min and the protein bands were then transferred to a single PVDF membrane at 1.0 A and 25 V for 30 min. The protein of interest and the internal control of protein loading, β-actin (43 kDa), were separated by cutting the membrane at around 50 kDa using the molecular weight marker as reference. The separated membranes were then placed in different containers and incubated with respective antibodies. After immunoblotting, the separated membranes were placed together and visualized using ChemiDoc TM XRS Imaging System (Bio-Rad Laboratories Inc, USA). For ( B ) hERG1a β-actin, the exposure time was 24 s, meanwhile for hERG1b β-actin, the exposure time was 111 s. As for ( D ) hERG1a β-actin, the exposure time was 16 s, meanwhile, for hERG1b and β-actin, the exposure time were 206 s and 32 s respectively. Densitometric scanning analyses for the protein expression of hERG1a (155 kDa and 135 kDa) and hERG1b (95 kDa and 80 kDa) are normalized against actin and relative to the untreated control. Densitometric data are presented as mean fold change ± SD of three independent experiments. *p
Article Snippet: Chemiluminescence signal was detected using ChemiDoc™ XRS Imaging System (Bio-Rad Laboratories, USA).
Techniques: Expressing, Western Blot, Electrophoresis, Molecular Weight, Marker, Incubation, Imaging