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·756 · 南 京 医 科 大 学 学 报 2024年6月

径降解，利用蛋白酶体抑制剂MG132（20 μmol/L）处 表明在Senp1 MEFs中Spop通过蛋白酶体途径降解。
-/-
理 Senp1 -/- MEFs。WB 结果显示，当用 MG132 抑制 2.3 Senp1不影响Spop的亚细胞定位
蛋白酶体活性后，细胞中的 Spop 有所升高（图 2C）， SUMO 化修饰除影响底物蛋白的周转速率外，

A B 1.5
Relative Spop protein level
WT Senp1 -/- ***

Spop 1.0
- 40 kDa 0.5

α⁃Tubulin - 55 kDa
0
WT Senp1 -/-
C 1.5 D
WT Senp1 -/-
mRNA level 1.0 Empty vector + - + - - -
Myc⁃Senp1

Spop 0.5 Myc - 70 kDa
Relative Spop - 40 kDa

0 α⁃Tubulin - 55 kDa
WT Senp1 -/-
A：Spop protein levels in WT and Senp1 -/- MEFs were measured by Western blot. B：Western blot results were quantified using Image J. C：Spop
mRNA levels in WT and Senp1 -/- MEFs were measured by qPCR. D：Changes in Spop protein levels were detected after gradient overexpressing of
-/-
Senp1 in Senp1 MEFs. Red arrows indicated Spop proteins. *** P < 0.001（n=3）。
图1 敲除Senp1下调Spop蛋白水平，但不影响其mRNA水平
Figure 1 Senp1 knockout down⁃regulated Spop protein levels，but does not affect mRNA levels

A CHX 0 h 2 h 3 h 4 h 5 h 6 h
Spop
WT - 40 kDa
α⁃Tubulin - 55 kDa

Spop
Senp1 -/- - 40 kDa

α⁃Tubulin - 55 kDa
B 1.00 Senp1 -/- C WT + Senp1 -/- +
Relative Spop protein level 0.75 MG132 - - + - 40 kDa
WT
+
DMSO
0.50
Spop
0.25

0 α⁃Tubulin - 55 kDa
0 h 2 h 4 h 6 h CHX
A：After the cells were treated with CHX（20 μmol/L）for different times，the protein levels of Spop in the cells were determined by Western blot.
B：Image J was used to normalize the results of panel A after quantification. C：After the cells were treated with MG132（20 μmol/L），the protein level of
Spop in the cells was determined by Western blot. Red arrows indicated Spop proteins.
图2 Senp1敲除后Spop蛋白稳定性下降
Figure 2 The stability of Spop protein decreased after Senp1 knockdown