Fig. 5: Lsm12 mediates the apparent association of NAADP to TPC2 and cell membranes. | Nature Communications

Fig. 5: Lsm12 mediates the apparent association of NAADP to TPC2 and cell membranes.

From: Lsm12 is an NAADP receptor and a two-pore channel regulatory protein required for calcium mobilization from acidic organelles

Fig. 5

a Immunoblot of TPC1 and TPC2 pulled down by immobilized NAADP from TPC1- and TPC2-expressing HEK293 WT and Lsm12-KO cells. b Immunoblot of Lsm12 pulled down by immobilized NAADP from cells with/without exogenous TPC1 and TPC2 expression. c Competition radioligand binding assay of the association between 32P-NAADP and TPC2-expressing cell membranes in the presence of various concentrations of non-radiolabeled NAADP. d Specific binding of 32P-NAADP to cell membranes in WT and Lsm12-KO cells with/without exogenous TPC2 expression. e In situ PLA of TPC2 and Lsm12 colocalization in HEK293 cells. Top panels show cells expressing both exogenous TPC2 and Lsm12 and bottom panels show cells expressing only one of them. AP affinity precipitation, Exp expression, AP: NAADP± AP with/without immobilized NAADP. Data were presented as mean value ± SEM. Unpaired Student’s t-test (two-tailed) was used to calculate p values. * is for p values ≤ 0.05.

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