Journal: American Journal of Physiology - Renal Physiology

Article Title: Effect of renal tubule-specific knockdown of the Na + /H + exchanger NHE3 in Akita diabetic mice

doi: 10.1152/ajprenal.00497.2018

Figure Lengend Snippet: Real-time PCR primers used

Article Snippet: For some genes and proteins, whole kidney expression was also estimated by considering kidney weight because diabetes had a strong effect on the latter. table ft1 table-wrap mode="anchored" t5 Table 1. caption a7 Target Gene Assay ID Actb Mm00607939_s1 Atp6v1b Mm00460309_m1 Ccl2 Mm00441242_m1 Cd14 Mm00438094_g1 Glud1 Mm00492353_m1 Glut1 Mm00441480_m1 Glut2 Mm00446229_m1 G6pc Mm00839363_m1 Nhe3 Mm01352473_m1 Pepck Mm01247058_m1 Pfkp Mm00444792_m1 Pkm2 Mm00834102_gH Renin Mm02342889_g1 Rhcg Mm00451199_m1 Rpl19 Mm02601633_g1 Sglt1 Mm00451203_m1 Tnfa Mm00443258_m1 Txnip Mm01265659_g1 Open in a separate window See text for the definitions of the abbreviations.

Techniques: Real-time Polymerase Chain Reaction, Gene Assay

Journal: American Journal of Physiology - Renal Physiology

Article Title: Effect of renal tubule-specific knockdown of the Na + /H + exchanger NHE3 in Akita diabetic mice

doi: 10.1152/ajprenal.00497.2018

Figure Lengend Snippet: Compensatory changes in renal gene expression in nondiabetic and diabetic Na+/H+ exchanger isoform 3 (NHE3) knockdown [NHE3-knockout (KO)] mice. Renal mRNA expression normalized to β-actin and ribosomal protein 119 (rpl19) is shown (n = 9–10 mice/group). A: NHE3-KO increased renal mRNA expression of genes associated with ammonium formation [glutaminase (Gls) and glutamate dehydrogenase (Glud1)], H+ secretion [H+-ATPase B1 subunit (Atp6v1b1)], and ammonia secretion [Rhesus C glycoprotein (Rhcg)]. These effects were more apparent in Akita diabetic mice. WT, wild type. B: both NHE3-KO and Akita diabetes increased renal phosphoenolpyruvate carboxykinase (Pepck) mRNA expression. C: in nondiabetic and diabetic mice, NHE3-KO enhanced renal mRNA expression of phosphofructokinase platelet isoform (Pfkp), pyruvate kinase M2 (Pkm2), and pyruvate kinase liver/red blood cell isoform (Pklr), which are rate-limiting enzymes of glycolysis. Akita diabetes also increased Pfkp and Pkm2. Values are means ± SE. Two-way ANOVA was performed to probe for a significant effect of NHE3-KO (PNHE3), Akita diabetes (PAkita), or the interaction between the two factors (Pinter). If the interaction was statistically significant, then a pair-wise multiple comparison procedure (Holm-Sidak method) identified the significant effects.

Article Snippet: For some genes and proteins, whole kidney expression was also estimated by considering kidney weight because diabetes had a strong effect on the latter. table ft1 table-wrap mode="anchored" t5 Table 1. caption a7 Target Gene Assay ID Actb Mm00607939_s1 Atp6v1b Mm00460309_m1 Ccl2 Mm00441242_m1 Cd14 Mm00438094_g1 Glud1 Mm00492353_m1 Glut1 Mm00441480_m1 Glut2 Mm00446229_m1 G6pc Mm00839363_m1 Nhe3 Mm01352473_m1 Pepck Mm01247058_m1 Pfkp Mm00444792_m1 Pkm2 Mm00834102_gH Renin Mm02342889_g1 Rhcg Mm00451199_m1 Rpl19 Mm02601633_g1 Sglt1 Mm00451203_m1 Tnfa Mm00443258_m1 Txnip Mm01265659_g1 Open in a separate window See text for the definitions of the abbreviations.

Techniques: Expressing, Knock-Out

Journal: Nature

Article Title: A single cell atlas of the tracheal epithelium reveals the CFTR-rich pulmonary ionocyte

doi: 10.1038/s41586-018-0394-6

Figure Lengend Snippet: a , Immunofluorescence for FOXI1 (red, arrow), and airway lineage markers (green, arrowheads); TP63 (basal), FOXJ1 (ciliated), MUC5B (secretory) and ASCL1 (PNEC) in differentiated HBEC cultures (n=3 donors). b , Fluorescent in situ hybridization (RNAscope ® ) in mouse tracheal epithelium (n=3 mice) and human bronchial epithelium (n=2 donors) for FOXI1 (red) and CFTR (green). c , HBECs were transduced at seeding with GFP or GFP:FOXI1 lentivirus, differentiated and then profiled by scRNAseq or analyzed by immunofluorescence (IF). d , Immunofluorescence for ATP6V1B1 (white) and FOXI1 (red) in HBECs transduced with GFP or GFP:FOXI1 (n=4 experiments from two donors). Scale bars, 20μm. e,f , Fold-change in fractions of cell states revealed by scRNA-seq in GFP:FOXI1 vs. GFP. Heatmap values correspond to the ratio of cell numbers from the viral transduction experiments projecting onto each point of the reference HBEC data set from . extends to populations specific to viral transduction. g , Ionocytes induced by GFP:FOXI1 are transcriptionally similar to natural ionocytes, shown by comparing their gene expression in scRNA-seq data from three experimental conditions (Reference data from ). The genes shown are markers of each epithelial cell type (bottom), with ionocyte markers shown in detail (top). Genes are normalized to the median expression level across populations observed in a given condition.

Article Snippet: Taqman probes for qPCR (Applied Biosystems) are shown below: FOXI1 , Hs00201827_m1 FOXJ1 , Hs00230964_m1; P63 , Hs00978340_m1; GAPDH , Hs99999905_m1; CFTR , Hs00357011_m1; ATP6V1B1 , Hs00266092_m1; ITGA6 , Hs01041011_m1; DNAI2 , Hs01001544_m1; SCGB1A1 , Hs00171092; MUC5B , Hs00861588_m1; NRARP , Hs01104102_s1; HES5 , Hs01387464_g1; HES1 , Hs00172878_m1; MUC5AC , Hs01365601_m1

Techniques: Immunofluorescence, In Situ Hybridization, RNAscope, Transduction, Gene Expression, Expressing