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, Control

Journal: Journal of Biological Chemistry

Article Title: Effect of Human Carbonic Anhydrase II on the Activity of the Human Electrogenic Na/HCO3 Cotransporter NBCe1-A in Xenopus Oocytes

doi: 10.1074/jbc.m602181200

Figure Lengend Snippet: FIGURE 6. Immunolocalization of EGFP and CA II domains in oocytes expressing EGFP-e1-CAII or EGFP- e1. Three days after injection of cRNA encoding EGFP-e1-CAII or EGFP-e1, oocytes were prepared for immu- nocytochemistryasindicatedunder“ExperimentalProcedures.”Preparationswereincubatedovernightsimul- taneously with mouse monoclonal anti-EGFP and with rabbit polyclonal anti-human CA II. The secondary antibodies were Alexa Fluor 488-conjugated goat anti-mouse and Alexa Fluor 594-conjugated goat anti-rab- bit. A-C, Alexa Fluor 488-labeled images were taken with excitation/emission filters set at band pass 488 nm/long pass 505 nm to detect EGFP signals. D-F, Alexa Fluor 594-labeled images were taken with excitation/ emission filters set at band pass 543 nm/long pass 560 nm to detect CA II signals. These images show that EGFP-e1-CAIIandEGFP-e1arecorrectlytargetedtotheplasmamembrane.G-I,mergesoftheimagesinthefirst and second columns. G shows that the EGFP staining (green) of EGFP-e1-CAII overlapped with the CA II staining (red). J–L, Nomarski images taken simultaneously with either the EGFP or CA II images. Bar 50 m.

Article Snippet: Slides were blocked by washing (once for 15 min) in TBS 0.1% BSA 10% normal goat serum (Vector Laboratories, Burlingame, CA).We incubated the preparations with mouse monoclonal anti-EGFP (BD Biosciences; 1:100 in TBS 0.1% BSA 10% goat serum) and rabbit polyclonal anti-human CA II (Rockland, Gilbertville, PA; 1:1000 in TBS 0.1% BSA 10% goat serum) overnight.

Techniques: Expressing, Injection, Labeling, Staining