Journal: American Journal of Physiology - Renal Physiology

Article Title: Effects of extreme potassium stress on blood pressure and renal tubular sodium transport

doi: 10.1152/ajprenal.00527.2019

Figure Lengend Snippet: Dietary K+ maneuvers affect the abundance of transporters and channels along the nephron. Kidney lysates from mice subjected to various dietary K+ maneuvers for 10 days were assessed by Western blot analysis. Equivalent protein loading was determined by BCA Protein Assay and Coomassie staining. Results are shown as means ± SE; n = 6 mice per diet. One-way ANOVA with Sidak’s post hoc analysis, *P ≤ 0.05 and **P ≤ 0.01 compared with control; φsignificant difference between K+ basic and KCl diets. A: we probed for a variety of channels and transporters expressed from the proximal tubule to the collecting duct, as indicated. B: Western blots of kidney homogenates from mice subjected to various K+ diets. All samples were of the kidney cortex unless noted [Na+-K+-2Cl− cotransporter (NKCC2) in the whole kidney]. #Multimerization of Na+-bicarbonate cotransporter 1A (NBCe1A) (39) and pendrin (62). C: graphical summary of the results. The dotted line depicts control values normalized to 1.0. DCT, distal convoluted tubule; γ-ENaC, γ-subunit of the epithelial Na+ channel; IC, intercalated cells; NCC, NaCl cotransporter; NHE3, Na+/H+ exchanger 3; ROMK, renal outer medullary K+ channel; NDCBE, Na+-driven Cl−/bicarbonate exchanger; PC, principal cells; PCT, proximal convoluted tubule; SGLT2, Na+-glucose transporter 2; TAL, thick ascending limb.

Article Snippet: The following antibodies were used for immunoblot analysis: aquaporin 2 (Aqp2; 29 kDa unglycosylated and 35−45 kDa mature, sc-515770, Santa Cruz Biotechnology) ( 46 ), Na + /H + exchanger 3 (NHE3; 84 kDa, SPC-400, Stressmarq) ( 28 ), Na + -bicarbonate cotransporter 1A (NBCe1A; 130 kDa denatured and 280 kDa dimer, AB3212-I, Millipore) ( 39 , 49 ), Na + -glucose transporter 2 (SGLT2; 74 kDa, ab37296, Abcam) ( 44 ), Na + -K + -2Cl − cotransporter (NKCC2; 160 kDa, SPC-401, Stressmarq) ( 6 ), NaCl cotransporter (NCC; 130 kDa, kindly provided by David Ellison) ( 8 ), phosphorylated (p)NCC (Thr 53 ; 130 kDa, P1311-53, Phospho-solutions) ( 40 ), γ-subunit of the epithelial Na + channel (γ-ENaC; 83 kDa cleaved and 95 kDa uncleaved, SPC-405, Stressmarq) ( 6 ), renal outer medullary K + channel (ROMK; 50−65 kDa complex glycosylated, R-80, kindly provided by James Wade) ( 61 ), electroneutral Cl − exchanger (pendrin; 85−110 kDa denatured and 220 kDa dimer, sc16894, Santa Cruz Biotechnology) ( 25 , 62 ), and Na + -driven Cl − /bicarbonate exchanger (NDCBE; 123 kDa, no. 12531-1, ProteinTech) ( 53 ).

Techniques: Western Blot, Bicinchoninic Acid Protein Assay, Staining

Journal: Journal of Interferon & Cytokine Research

Article Title: IFN-α-Induced Downregulation of miR-221 in Dendritic Cells: Implications for HCV Pathogenesis and Treatment

doi: 10.1089/jir.2014.0211

Figure Lengend Snippet: Interferon (IFN)-α induces downregulation of miR-221-3p in dendritic cells (DCs). (A) Heat map representing mean fold change in expression of 113 miRNAs (84 miRNAs profiled using Human miFinder miScript miRNA qPCR Array, SA Biosciences plus 29 miRNAs whose qPCR primers were designed according to similar strategy as miFinder array) in myeloid dendritic cells (mDCs) (n=2) upon 24 h IFN-α treatment. Asterisk (*) indicates miRNA with greater than 1.5-fold upregulation/downregulation in both biological replicates. (B) Bar graph indicating mean fold change in expression of 7 miRNAs (which were marked with asterisk in the previous graph) in mDCs (n=3). (C) Bar graph indicating mean fold change in expression of same 7 miRNAs in plasmacytoid dendritic cells (pDCs) (n=3) upon 24 h IFN-α treatment. (D) Out of the remaining 116 miRNAs profiled in pDCs, 16 miRNAs showed significant downregulation upon 24 h IFN-α treatment. Bar graph indicates the mean fold-downregulation of those 16 miRNAs. (E) Bar graph indicating mean fold change in expression of miR-221 in B cells upon 24 h IFN-α treatment. (F) Bar graph indicating the kinetics of IFN-α-induced downregulation of miR-221 in mDCs. P values were calculated using Student's t test (*P<0.05; **P<0.01). Error bars represent standard deviation (n=3). Color images available online at www.liebertpub.com/jir

Article Snippet: In subsequent experiments, we decided to focus on the mechanism of miR-221 downregulation and its impact on mDCs. fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window FIG. 1. caption a7 Interferon (IFN)-α induces downregulation of miR-221-3p in dendritic cells (DCs). (A) Heat map representing mean fold change in expression of 113 miRNAs (84 miRNAs profiled using Human miFinder miScript miRNA qPCR Array, SA Biosciences plus 29 miRNAs whose qPCR primers were designed according to similar strategy as miFinder array) in myeloid dendritic cells (mDCs) ( n =2) upon 24 h IFN-α treatment.

Techniques: Expressing, Standard Deviation

Journal: Toxins

Article Title: Bacillus thuringiensis Bt_UNVM-84, a Novel Strain Showing Insecticidal Activity against Anthonomus grandis Boheman (Coleoptera: Curculionidae)

doi: 10.3390/toxins16010004

Figure Lengend Snippet: Detection of cry genes and proteins from the parasporal crystals. ( A ) Detection of cry genes by PCR with degenerate primers . Amplicons were electrophoresed in 1% agarose gel: M molecular weight marker 1 kb, (+) B. thuringiensis strain HD1 control, (−) negative control with water; 81 is a negative control strain whereas 84 is the new isolate and ( B ) SDS-PAGE analysis: M molecular weight marker (Precision Plus Proteins Dual Color), 84 dried Bt_UNVM-84 biomass, 84 1 solubilized Bt_UNVM-84 biomass, and 84 2 solubilized Bt_UNVM-84 biomass digested (potentially activated) with the enzyme trypsin.

Article Snippet: Mapping analysis using Bt_UNVM-84 Illumina reads on the pFR260 plasmid sequence showed 96.1% pairwise identity, covering 88.1% of the plasmid (used as the reference sequence).

Techniques: Agarose Gel Electrophoresis, Molecular Weight, Marker, Negative Control, SDS Page

Journal: Toxins

Article Title: Bacillus thuringiensis Bt_UNVM-84, a Novel Strain Showing Insecticidal Activity against Anthonomus grandis Boheman (Coleoptera: Curculionidae)

doi: 10.3390/toxins16010004

Figure Lengend Snippet: GBDP tree (whole-genome sequence-based) using TYGS server (average branch support 98.5%) . Green color was used to highlight Bt_UNVM-84 strain clustered along with type strains.

Article Snippet: Mapping analysis using Bt_UNVM-84 Illumina reads on the pFR260 plasmid sequence showed 96.1% pairwise identity, covering 88.1% of the plasmid (used as the reference sequence).

Techniques: Sequencing