Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

Article Title: Cellular chloride and bicarbonate retention alters intracellular pH regulation in Cftr KO crypt epithelium

doi: 10.1152/ajpgi.00236.2015

Figure Lengend Snippet: Gene name, symbol, and Life Technologies assay ID for custom TaqMan mini-array

Article Snippet: The threshold cycle (C t ) of a gene of interest was subtracted from the geometric mean C t of three housekeeping genes to yield ΔC t . The relative mRNA expression of Cftr KO relative to WT enteroids was calculated using the ΔΔC t method ( 40 ). table ft1 table-wrap mode="anchored" t5 Table 1. caption a7 Gene Name Symbol Assay ID Glucuronidase, β * Gusb Mm00446953_m1 Hypoxanthine guanine phosphoribosyl transferase 1 * Hprt1 Mm00446968_m1 Mitochondrial ribosomal protein L19 * Mrpl19 Mm00452754_m1 Sodium/potassium ATPase-α 1 (Na + -K + -ATPase) Atp1a1 Mm00523255_m1 Carbonic anhydrase II ( CA II ) Car2 Mm00501572_m1 Carbonic anhydrase IX ( CA IX ) Car9 Mm00519870_m1 Sodium-potassium-2 chloride cotransporter 1 ( Nkcc1 ) Slc12a2 Mm00436554_m1 Downregulated in adenoma ( Dra ) Slc26a3 Mm00445313_m1 Putative anion transporter 1 ( Pat-1 ) Slc26a6 Mm00506742_m1 Anion exchanger 2 ( Ae2 ) Slc4a2 Mm00436617_m1 Sodium-bicarbonate cotransporter e1 (Nbce1) Slc4a4 Mm01347935_m1 Sodium/hydrogen exchanger 1 ( Nhe1 ) Slc9a1 Mm00444270_m1 Sodium/hydrogen exchanger 2 ( Nhe2 ) Slc9a2 Mm01237129_m1 Sodium/hydrogen exchanger 3 ( Nhe3 ) Slc9a3 Mm01352473_m1 Anoctamin 1 ( Ano1 ) Ano1 Mm00724407_m1 Open in a separate window * Housekeeping gene.

Techniques:

Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

Article Title: Cellular chloride and bicarbonate retention alters intracellular pH regulation in Cftr KO crypt epithelium

doi: 10.1152/ajpgi.00236.2015

Figure Lengend Snippet: Changes in expression of pHi regulators in Cftr KO crypt enteroids. A: quantitative real-time PCR of mRNA from 8 sex-matched WT and Cftr KO littermate pairs. Genes represented are the Na+-K+-ATPase, carbonic anhydrases (CA II and CA IX), Na+-K+-2Cl− cotransporter (Nkcc1, Slc12Aa2), downregulated in adenoma (Dra, Slc26a3), putative anion transporter-1 (Pat-1, Slc26a6), anion exchanger 2 (Ae2, Slc4a2), electrogenic NaHCO3 cotransporter 1 (Nbce1, Slc4a4), Na+/H+ exchangers 1, 2, and 3 (Nhe1-3, Slc9a1-3), and the anion channel anoctamin 1 (Ano1, Tmem16a). *Significantly different from 0 (by 1-sample t-test). B: representative immunoblot for Ae2 and β-actin of protein lysate from enteroids cultured from a sex-matched WT and Cftr KO littermate pair and a sex-matched Ae2 KO mouse. C: cumulative data for Ae2 immunoblot densitometry of enteroids from sex-matched WT and Cftr KO littermate pairs. Values are means ± SE (n = 11). *Significantly different from WT (by unpaired t-test). D: cumulative data for early growth response-1 (Egr-1) immunoblot densitometry of enteroids from sex-matched WT and Cftr KO littermate pairs. Values are means ± SE (n = 8). *Significantly different from WT (by unpaired t-test).

Article Snippet: The threshold cycle (C t ) of a gene of interest was subtracted from the geometric mean C t of three housekeeping genes to yield ΔC t . The relative mRNA expression of Cftr KO relative to WT enteroids was calculated using the ΔΔC t method ( 40 ). table ft1 table-wrap mode="anchored" t5 Table 1. caption a7 Gene Name Symbol Assay ID Glucuronidase, β * Gusb Mm00446953_m1 Hypoxanthine guanine phosphoribosyl transferase 1 * Hprt1 Mm00446968_m1 Mitochondrial ribosomal protein L19 * Mrpl19 Mm00452754_m1 Sodium/potassium ATPase-α 1 (Na + -K + -ATPase) Atp1a1 Mm00523255_m1 Carbonic anhydrase II ( CA II ) Car2 Mm00501572_m1 Carbonic anhydrase IX ( CA IX ) Car9 Mm00519870_m1 Sodium-potassium-2 chloride cotransporter 1 ( Nkcc1 ) Slc12a2 Mm00436554_m1 Downregulated in adenoma ( Dra ) Slc26a3 Mm00445313_m1 Putative anion transporter 1 ( Pat-1 ) Slc26a6 Mm00506742_m1 Anion exchanger 2 ( Ae2 ) Slc4a2 Mm00436617_m1 Sodium-bicarbonate cotransporter e1 (Nbce1) Slc4a4 Mm01347935_m1 Sodium/hydrogen exchanger 1 ( Nhe1 ) Slc9a1 Mm00444270_m1 Sodium/hydrogen exchanger 2 ( Nhe2 ) Slc9a2 Mm01237129_m1 Sodium/hydrogen exchanger 3 ( Nhe3 ) Slc9a3 Mm01352473_m1 Anoctamin 1 ( Ano1 ) Ano1 Mm00724407_m1 Open in a separate window * Housekeeping gene.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Western Blot, Cell Culture