Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Identification of Novel Laminin- and Fibronectin-binding Proteins by Far-Western Blot: Capturing the Adhesins of Streptococcus suis Type 2

doi: 10.3389/fcimb.2015.00082

Figure Lengend Snippet: Determine the binding of the recombinant proteins to LN and FN by Far-Western blot . Coomassie G-250-stained gel (A) , Western blot analysis (B) , and Far-Western blot analysis (C,D) of the SS2 recombinant proteins. Recombinant proteins were separated by 12% SDS-PAGE, then transferred to PVDF membrane and incubated with human LN or LN. Bound LN or FN was detected with goat anti-rabbit IgG antibody. The Western blot (B) was probed with his tag monoclonal antibody (Boster).

Article Snippet: His tag monoclonal antibody (Boster; 1:1000 dilution) were added to every plate and wells were then incubated at 37°C for 30 min. After washing, the cultured cells were incubated with goat anti-mouse IgG-FITC (Boster; 1:2000 dilution) at 37°C for 30 min. After a finial wash, the samples were examined using a fluorescence microscope (ZEISS, Germany).

Techniques: Binding Assay, Recombinant, Far Western Blot, Staining, Western Blot, SDS Page, Membrane, Incubation

Journal: Molecular Medicine Reports

Article Title: Reoxygenation induces reactive oxygen species production and ferroptosis in renal tubular epithelial cells by activating aryl hydrocarbon receptor

doi: 10.3892/mmr.2020.11679

Figure Lengend Snippet: AhR activation status does not affect Nrf2 activation or transcriptional activity. RPTECs were cultured under ctrl conditions or subjected to Reox with or without the AhR inhibitor CH223191. (A) Representative western blots of Nrf2 levels (corresponding to its activation status) and the expression of the Nrf2 transcriptional targets xCT (SLC7A11) and SOD-3. (B-D) Statistical analysis of the western blots. Neither Reox nor CH223191 affects Nrf2 activity, or the expression of xCT and SOD-3. Data are presented as the mean ± SEM of six independent experiments. AhR, arylhydrocarbon receptor; Nrf2, nuclear factor erythroid 2-related factor 2; xCT, cystine-glutamate antiporter; SOD-3, superoxide dismutase; RPTEC, renal proximal tubular epithelial cell; ctrl, control; Reox, reoxygenation.

Article Snippet: Primary antibodies were specific for AhR (1:200; cat. no. sc-133088; Santa Cruz Biotechnology, Inc.), cytochrome P450 family 1 subfamily A member 1 (CYP1A1; 1:500; cat. no. sc-25304; Santa Cruz Biotechnology, Inc.), Nrf2 (1:1,000; cat. no. TA343586; OriGene Technologies, Inc.), superoxide dismutase 3 (SOD-3; 1:100; cat. no. sc-271170; Santa Cruz Biotechnology, Inc.), cystine-glutamate antiporter (xCT, also known as SLC7A11; 1:1,000; cat. no. ANT-111; Alomone Labs), HIF-1α (1:500; cat. no. sc-10790; Santa Cruz Biotechnology, Inc.), LDH-A (1:1,000; cat. no. 2012; Cell Signaling Technology, Inc.), activated cleaved caspase-3 (CC3; 1:1,000; cat. no. ab13847; Abcam) and β-actin (1:2,500; cat. no. 4967; Cell Signaling Technology, Inc.).

Techniques: Activation Assay, Activity Assay, Cell Culture, Western Blot, Expressing