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Supplement Figure 1: The expression of SIRT4 in some other cancers.
SIRT4 was positive in normal tissues comparing with corresponding
tumor tissues.
A, normal ovarian tissues. B, ovarian cancer tissues. C, normal liver
tissues. D, liver cancer tissues. E normal prostate tissues. F prostate
cancer tissues. G normal pancreatic tissues. H pancreatic cancer tissues.I
normal bladder tissues. J,bladder cancer tissues K normal renal tissues. L,
renal cancer tissues.
Supplement Figure 2: 3 siRNA sequences from Dharmacon and used
siRNA mix to reduce the risk of off target effects. The inference
efficiency was shown by wstern blot assay and quantified with relative
grey-value. The mixed use of the three sequences made a significant
reduce of endogenous SIRT4 expression.
Supplement Figure 3: Cells were collected for western blot assay at 2d,
4d, 6d, 10d, 12d and 16d after transfection. The timeline of tranfection
efficiency was quantified by relative grey-value. From the results, we
believe that the transfection efficiency lasted for at least 1 week, which
was verified by the western blot assay.
Supplement Figure 4: Fluorescence staining was performed to show the
homogeneous overexpression of SIRT4 in all H1299 and A549 cells
respectively after SIRT4 plasmids transfection, comparing with
corresponding empty vector.