shrna expressing mcf 7 cells  (Santa Cruz Biotechnology)

Journal: Science signaling

Article Title: Plasma membrane Ca²⁺-ATPases can shape the pattern of Ca²⁺ transients induced by store-operated Ca²⁺ entry.

doi: 10.1126/scisignal.2005672

Figure Lengend Snippet: Fig. 1. Generation and characterization of a PMCA4 knockdown HeLa cell line. (A) PMCA abundance in control HeLa (nontransfected) and sh-PMCA4 (sh4) HeLa cells grown for 3, 5, or 7 days. Western blot analysis was per- formed with equal amounts (20 mg) of whole-cell lysates using either the 5F10 antibody to detect both PMCA1 and PMCA4 (top) or JA3 antibody specific for PMCA4b (middle). Na+,K+-ATPase served as a loading control. (B) Representative experiment of the Ca2+ re-addition protocol for studying SOCE. The recordings are from sh-PMCA4 HeLa cells. Thapsigargin (Tg) inhibits SERCA, ATP stimulates release from the ER store, and addition of 2 mM Ca2+ initiates SOCE. (C) SOCE-mediated Ca2+ transients in individ- ual control HeLa cells expressing GCaMP2. (D) SOCE-mediated Ca2+

Article Snippet: The sh-PMCA4 HeLa cell line was generated by transfecting HeLa cells with a PMCA4 shRNA plasmid (Santa Cruz Biotechnology Inc.) as described (29).

Techniques: Knockdown, Control, Western Blot, Expressing

Journal: Science signaling

Article Title: Plasma membrane Ca²⁺-ATPases can shape the pattern of Ca²⁺ transients induced by store-operated Ca²⁺ entry.

doi: 10.1126/scisignal.2005672

Figure Lengend Snippet: Fig. 5. PMCA isoforms differentially affect SOCE induced at different extra- cellular Ca2+ concentrations. (A to D) After store depletion, SOCE was ini- tiated in HeLa cells expressing the indicated mCherry-PMCA constructs or in sh-PMCA4 HeLa cells by the stepwise addition of increasing [Ca2+]e, and intracellular [Ca2+]i was followed for 5 min after each addition. Average traces from about 20 cells (or 8 cells for PMCA4b-LA) are shown (means ± 95% CI). Ca2+ signals in all panels were detected by the fluorescence of GCaMP2.

Article Snippet: The sh-PMCA4 HeLa cell line was generated by transfecting HeLa cells with a PMCA4 shRNA plasmid (Santa Cruz Biotechnology Inc.) as described (29).

Techniques: Expressing, Construct, Fluorescence