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. 2011 Apr 12;286(23):20194–20207. doi: 10.1074/jbc.M110.148494

FIGURE 3.

FIGURE 3.

Silencing TRPM7 with shRNA down-regulates the expression of TRPM7. A shows the infection efficiency of lentivirus carrying shRNA-EGFP/TRPM7 and shRNA-EGFP/Control. Hoechst 33258 was used to dye the cell nucleus. shRNA/TRPM7 lentivirus could infect ∼60% neurons, whereas shRNA/control infected ∼80%. The bar in the right corner represents 100 μm. B shows the specificity of TRPM7 antibody. The upper and lower images represent cells stained with goat polyclonal TRPM7 antibody and normal goat IgG, respectively. The bar in the right corner represents 25 μm. C shows the effect of shRNA on TRPM7 expression in neurons with immunocytofluorescence. The bar in the right corner represents 50 μm. D shows the effect of shRNA on TRPM7 in mRNA levels using RT-PCR (averaged from five independent experiments). The mRNA of TRPM7 was reduced by 53.61 ± 2.53% using shRNA aimed at TRPM7. * represents having significant difference (p < 0.05) compared with the normal group. E shows the effect of shRNA on TRPM7 in protein level using Western blotting (averaged from three independent experiments). TRPM7 expression was reduced by 42.67 ± 5.03% shRNA aimed at TRPM7. * represents having significant difference (p < 0.05) compared with the normal group.