Fig. 1.Knockdown of TRPM7 in DFCs using lentiviruses that express
TRPM7-specific shRNA. (a) TRPM7 mRNA, but not TRPM6 mRNA, was detected in DFCs
by reverse transcription PCR. CHO cells with ectopic expression of TRPM6 or TRPM7
were used as positive controls. (b) qRT-PCR showed that TRPM7 gene expression was
significantly reduced in TRPM7 knockdown cells. Quantitative data from three
independent experiments is shown. **, p 0.01. (c) Western blot was
performed to detect the TRPM7 protein expression. Proteins isolated from CHO
cells transduced with TRPM7 served as positive controls. lanes 1 and 2: CHO cells
transduced with TRPM7. lanes 3 and 4: DFCs transduced with scrambled shRNA. lanes
5 and 6: DFCs transduced with TRPM7 shRNA. (d) Immunocytochemistry showed that
TRPM7 was highly expressed in DFCs, but barely expressed in TRPM7 knockdown
cells. Scale bar: 100 M. (e) Typical whole cell TRPM7-like currents
recorded in DFCs in the absence of Mg (black color), in the presence of 2
mM 2APB (red color), of 100 M 2APB (pink color), or infected with
TRPM7-specific shRNA (blue color).