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. Author manuscript; available in PMC: 2018 Aug 26.
Published in final edited form as: Oncogene. 2018 Feb 26;37(20):2615–2629. doi: 10.1038/s41388-018-0138-y

Figure 7. UGDH is required for KLF4-mCpG-dependent increase in GBM cell migration.

Figure 7

(A) U87 KLF4 WT and GBM1A KLF4 WT cells were transduced with lentivirus coding for nonsilencing shRNA (Control), UGDH shRNA #1 or UGDH shRNA #2. Cells were treated with Dox for 48 hrs before each analysis. Western blot showed UGDH knockdown significantly reverses KLF4 WT dependent induction of UGDH in both U87 and GBM1A cells. (B) Sulfated GAG (sGAG) concentration in U87 and GBM1A cells expressing KLF4 WT was determined by DMMB assay. UGDH knockdown significantly decreased the KLF4 dependent increase in GAG concentration (C) UGDH knockdown significantly reversed the KLF4-dependent increase in cell migration in transwell assays. Cell migration was evaluated 24 hrs later by counting DAPI-stained cells. Five fields per well were counted. (D) U87 KLF4 WT cells harboring UGDH shRNA were treated with Dox for 5 days till confluence. A scratch was made and cells were maintained in 0.1% FCS medium overnight. Microphotographs were taken 0 hr and 24 hrs after scratching. Bar = 100 um. UGDH knockdown inhibited the increased ability of KLF4 WT cells to migrate towards scratched area.