Journal: International Journal of Molecular Sciences

Article Title: Perineural Dexmedetomidine Attenuates Inflammation in Rat Sciatic Nerve via the NF-κB Pathway

doi: 10.3390/ijms15034049

Figure Lengend Snippet: Perineural administration of 20 μg/kg dexmedetomidine decreased the translocation to the nucleus and transcriptional binding activity of NF-κB. ( A ) Western blot showed that 20 μg/kg dexmedetomidine decreased NF-κB translocation into the nucleus; ( B ) Dexmedetomidine (20 μg/kg) decreased the nuclear NF-κB protein level; ( C ) Dexmedetomidine (20 μg/kg) did not alter the total NF-κB protein level; ( D ) EMSA showed that 20 μg/kg dexmedetomidine decreased the transcriptional binding activity of NF-κB (lane 3); PDTC also decreased NF-κB transcriptional binding activity (lane 5). NF-κB, nuclear factor-kappa B; PDTC, pyrrolidine dithiocarbamate.

Article Snippet: An electrophoretic-mobility shift assay (EMSA) kit (GS-0030, Signosis, Santa Clara, CA, USA) was used to assess the transcriptional binding activity of NF-κB as a previous study described [ ].

Techniques: Translocation Assay, Binding Assay, Activity Assay, Western Blot

Journal: Oncology Letters

Article Title: Differential behavior of NF-κB, IκBα and EGFR during the renal carcinogenic process in an experimental model in vivo

doi: 10.3892/ol.2020.11436

Figure Lengend Snippet: NF-κB activity analyzed by EMSA and EGFR mRNA levels determined by reverse transcription-PCR at early stages of FeNTA-induced renal carcinogenesis. Representative images of both determinations as well as corresponding histograms of densitometric analysis of all samples assayed are exhibited. (A) EMSAs were performed using nuclear extracts: Lane 1: Digoxigenin-labeled oligonucleotides without sample; lane 2: Sample without oligonucleotides; lane 3: Competition assay carried out using a 100-fold molar excess of unlabeled oligonucleotides; lane 4: Supershift assay performed utilizing an anti-p65 antibody. DF exposure induced an increase in NF-κB activity after one month, but it decreased after two months. (B) EGFR mRNA levels increased by a similar degree at both times of exposure. GAPDH was used as a loading control. Bars represent the mean ± standard error of the mean. n=4 for C groups and 7 for DF groups. *P≤0.05 vs. corresponding C and + P≤0.05 vs. DF after one month. C, control group; DF, N-dietilnitrosamine + FeNTA treated group; EMSA, electrophoretic mobility shift assay; RDU, relative densitometric units vs. GAPDH and with respect to the C group; NF-κB, nuclear factor kappa B; EGFR, epidermal growth factor receptor.

Article Snippet: Primary antibodies against IκBα (cat. no. sc-371), NF-κB p65 (cat. no. sc-8008), EGFR (cat. no. sc-03-G), α-tubulin (cat. no. sc-5286), GAPDH (cat. no. sc-48167), β-actin (cat. no. sc-1616) and histone H3 (cat. no. sc-10809) used for western blotting (WB), immunohistochemistry (IHC), and the supershift test of the electrophoretic mobility shift assay (EMSA), as well as the secondary antibodies (anti-mouse and anti-rabbit, cat. nos. sc-2005 and sc-2004, respectively) used for WB were purchased from Santa Cruz Biotechnology, Inc., while the anti-IgG secondary antibodies coupled with biotin (ABC kit Vectastain ® ; cat. nos.

Techniques: Activity Assay, Reverse Transcription, Labeling, Competitive Binding Assay, Control, Electrophoretic Mobility Shift Assay