Figure 8 (IMAGE)
Caption
NEAT1 plays a pivotal role in JunD-mediated downregulation of ZO-1. (A) Schematic representation of RNA immunoprecipitation (RIP) performed to determine the interaction between pJunD and NEAT1. (B) Expression of pJunD in absence and presence of 50 μM and 400 μM arecoline after immunoprecipitation with pJunD specific antibody. RIP with anti-IgG, indicating non-specific antibody binding, served as the negative control. (C) Expression of NEAT1 following RIP by semi-qPCR and RT-PCR. 18S served as control for non-specific amplification. (D) Expression of JunD and NEAT1 after silencing JunD in HEp-2 cells followed by treatment without and with 50 μM (+) and 100 μM (++) arecoline. The relative RNA expressions are represented for NEAT1 and JunD. (E) Expression of JunD and NEAT1 after silencing NEAT1 in HEp-2 cells followed by treatment without and with 50 μM (+) and 100 μM (++) arecoline. (F) Differential protein expression of tight junction and EMT markers in HEp-2 cells after silencing NEAT1 followed by treatment without and with 50 μM (+) and 100 μM (++) arecoline. β-tubulin was used as an internal control for all western blots. All the experiments were performed three times. Each value is the mean ± S.D. of three different replicate experiments, each performed in triplicate. *p < 0.1 and ***p < 0.001.
Credit
Correspondence to - Urmi Chatterji - urmichatterji@gmail.com
Usage Restrictions
Credit must be given to the creator.
License
CC BY