Skip to main content
. 2015 Apr 14;34(13):1743–1758. doi: 10.15252/embj.201490009

Figure 8.

Figure 8

TPC proteins are not necessary for NAADP binding
  • A RT–qPCR analysis of absolute levels of Tpcn1 and Tpcn2 transcripts in liver from WT animals. Tpcn1/Tpcn2 ratio of expression corresponds to 43.9; n = 6.
  • B RT–PCR analysis of Tpcn1 and Tpcn2 expression in wild-type (WT) or Tpcn1/2−/− (DKO) liver preparations. Expression of Actb was used as a control. Amplified cDNA regions correspond to the same exons as in Fig1D.
  • C, D [32P]NAADP-binding assay with competition by NAADP and NAADP-related dinucleotides performed with liver homogenates from WT (C, D) or DKO (D) animals. Data are expressed as values relative to total binding performed in the absence of unlabelled dinucleotide. (D) The IC50 values for the high-affinity binding site were WT: 13.7 ± 7.5 nM and DKO: 146.3 ± 60.5 nM (P > 0.09) and for the low-affinity site WT: 6.9 ± 4.1 μM and DKO: 27.4 ± 8.3 μM (P > 0.08); N (number of animals) = 5–7; n (number of binding reactions) = 10–14.
  • E Photoaffinity labelling of liver homogenates from WT or DKO animals performed with [32P]5N3-NAADP in the presence or absence of unlabelled NAADP (1 μM).
Data information: Error bars represent SEM. Source data are available online for this figure.