Journal: Nucleic Acids Research

Article Title: Functional transitions of the Aspergillus fumigatus iron regulator HapX are governed by conserved domains cooperatively binding [2Fe-2S] clusters

doi: 10.1093/nar/gkaf796

Figure Lengend Snippet: In liquid media, growth inhibition by combinatorial mutation of at least CRR-B and -C is more severe in +Fe compared to −Fe and accelerates with increased expression of the corresponding hapX alleles. For each strain, ( A ) pxylP:hapX , ( B ) pxylP:hapX A/B/C/D , ( C ) pxylP:hapX B/C , and ( D ) pxylP:hapX A/B/C/D,464 , 10 4 conidia were inoculated in triplicate into 96-well microplates in 0.1 mL minimal medium reflecting iron limitation (−Fe; in green) and iron sufficiency (+Fe; 10 μM FeSO 4 ; in red) supplemented with 0.1% xylose for moderate and 1% xylose for high expression of the respective hapX allele. Using the oCelloScope (BioSense Solutions, Denmark) and its UniExplorer software (version 14.0), growth was scored hourly at 37 °C from 2 h post-inoculation (graphs start at the 5 h time point as there was very limited growth before that) until 24 h incubation. In the software, the “ fungi filamentous growth module ” was used to assess the hyphal area up to 2.75 × 10 5 μm 2 when the growth measurement reached saturation. The values in the growth curves represent the mean ± standard deviation of biological triplicates. Below the growth curves are examples of microscopic images of the fungal cultures taken automatically by the oCelloScope at two time points, 12 h and 24 h, underlining the scored growth data.

Article Snippet: Therefore, we applied automated microbial live cell imaging and analysis using the oCelloScope (BioSense Solutions ApS, Denmark) measuring growth by determining hyphal area (Fig. ).

Techniques: Inhibition, Mutagenesis, Expressing, Software, Incubation, Standard Deviation

Journal: American Journal of Physiology - Heart and Circulatory Physiology

Article Title: The neuropilin-like protein ESDN regulates insulin signaling and sensitivity

doi: 10.1152/ajpheart.00782.2015

Figure Lengend Snippet: Adaptor protein with pleckstrin homology and Src homology 2 domain (APS)-IR interaction mediates the ESDN effect on insulin signaling. A: IR-β immunoprecipitation from the liver and muscle tissue extracts of WT and Esdn−/− mice in the absence of, and 50 min after, insulin (1 U/kg) administration, followed by immunoblotting for IR-β, ESDN, and ubiquitin. Data are representative of 3 independent experiments. B: IR-β immunoprecipitation from the liver and muscle tissue extracts of WT and Esdn−/− mice followed by immunoblotting for IR-β, APS, c-casitas B-lineage lymphoma (Cbl), growth factor receptor bound protein 10 (Grb10), and neuronal precursor cell-expressed developmentally downregulated 4 (Nedd4). Data from two representative pairs of animals are displayed. C: Western blot analysis of APS, c-Cbl, Grb10, and Nedd4 expression in the liver and muscle of WT and Esdn−/− mice, demonstrating a similar expression level. D: representative coimmunofluorescence staining of ESDN (in green, left) and APS or Grb10 (in red, middle) and fused images (right) demonstrating ESDN-APS and ESDN-Grb10 colocalization. Insets show lower-magnification whole cell images in each panel and fused control staining on right, where nuclei are stained blue with DAPI. Arrows in the insets point to the magnified cell surface area. Scale bar: 6 μm (12 μm for insets). E–H: representative Western blots (E and G) and quantification (F and H) of the effect of siRNA-mediated APS (E and F) or Grb10 (G and H) downregulation on insulin (105 nM, 20 min)-induced IR-β phosphorylation in WT and Esdn−/− VSMCS; n = 3. *P < 0.05, 2-way ANOVA.

Article Snippet: The antibodies for phospho-MAPK p44/42 (Thr 202/204 ), MAPK p44/42, phospho-Akt (Ser 473 ), Akt, phospho-IR-β (Tyr 1146 ), IR-β (L55B10 and 4B8), c-Cbl, APS, Grb10, Nedd4, ubiquitin, and GAPDH used for Western blotting were obtained from Cell Signaling Technologies (Danvers, MA).

Techniques: Immunoprecipitation, Western Blot, Ubiquitin Proteomics, Expressing, Staining, Control, Phospho-proteomics

Journal: American Journal of Physiology - Heart and Circulatory Physiology

Article Title: The neuropilin-like protein ESDN regulates insulin signaling and sensitivity

doi: 10.1152/ajpheart.00782.2015

Figure Lengend Snippet: Schematic presentation of the effect of ESDN on IR interaction with key regulatory molecules, APS, c-Cbl, Grb10, and Nedd4.

Article Snippet: The antibodies for phospho-MAPK p44/42 (Thr 202/204 ), MAPK p44/42, phospho-Akt (Ser 473 ), Akt, phospho-IR-β (Tyr 1146 ), IR-β (L55B10 and 4B8), c-Cbl, APS, Grb10, Nedd4, ubiquitin, and GAPDH used for Western blotting were obtained from Cell Signaling Technologies (Danvers, MA).

Techniques:

Journal: Frontiers in Pharmacology

Article Title: Purple corn extract improves benign prostatic hyperplasia by inhibiting 5 alpha-reductase type 2 and inflammation in testosterone propionate-induced rats

doi: 10.3389/fphar.2024.1485072

Figure Lengend Snippet: Effect of PCE on AR signaling-related protein expression in rats with TP-induced BPH. Immunohistochemical analyses were performed using anti-AR, anti-SRC-1, and anti-PSA antibodies. Representative images are presented in the original magnification ×60. Fina, finasteride; TP, testosterone propionate.

Article Snippet: Antibodies specific to IL-6 (MA5-45070), TNF-α (PA1-40281), and NRF-2 (PA1-38312) were obtained from Invitrogen (Waltham, MA, USA), and AR (ab133273), PSA (A01505-1), SRC-1 (sc-32789), and caspase-3 (NB100-56113) were purchased from Abcam (Cambridge, UK), Boster Biological Technology (Pleasanton, CA, USA), Santa Cruz Biotechnology (Santa Cruz, CA, USA), and Novus Biologicals (Centennial, CO, USA), respectively.

Techniques: Expressing, Immunohistochemical staining