Fig 1: Effects of temozolomide on regulation of skeleton and skeleton-related protein expression in GBM cells.(a) Rearrangement and damage of skeleton and immunofluorescence analysis of Actin, Tubulin, DHC2 and KIF2B in U87 cells after treatment with temozolomide/DMSO for two weeks. Magnification, ×1200. (b) Summarized data of immunofluorescence about these proteins (#P > 0.05; *P < 0.05; and **P < 0.01).
Fig 2: Expression of DHC2 and KIF2B in nude mouse tumor xenografts.(a) Tumors samples from xenograft model. (b) Immunohistochemistry analysis of DHC2 and KIF2B in tissue samples from xenograft models. (c) Expression of DHC2 in GBM tissue specimens. Immunohistochemical analysis of DHC2 in primary and recurrence GBM tissue samples.
Fig 3: Knockdown of DHC2 and KIF2B expression in U87 cells.(a) Western blot. Cells were grown and transfected with two different siRNAs for 8 h and cultured for 48 h and then subjected to Western blot analysis for protein expression. The graph is summarized data of the blot. *P < 0.05 and **P < 0.01. (b) Morphology and viability change of cells after knockdown of DHC2 and KIF2B expression followed by temozolomide treatment for one week. Cells were then counted and the graph shows the number of surviving cells in different groups (#P > 0.05; *P < 0.05 and **P < 0.01).
Fig 4: Knockdown of DHC2 expression in 2 different primary GBM cell lines.(a) Western blot. Cells were grown and transfected with two different siRNAs for 8 h and cultured for 48 h and then subjected to Western blot analysis of protein expressions. The graph is the summarized data of the blot. *P < 0.05; **P < 0.01. (b) Changes in viability of cells after knockdown of DHC2 expression followed by temozolomide treatment for one week. Cells were then counted and the graph showed the number of survival cells in different groups (#P > 0.05; *P < 0.05; and **P < 0.01).
Fig 5: qRT-PCR validation of DHC2 and KIF2B expression in temozolomide-treated cells.Expression of DHC2 and KIF2B mRNA was detected in GBM cells treated with 200 μM temozolomide or DMSO for one to two weeks using qRT-PCR (*P < 0.05 and **P < 0.01).
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