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Fig. 3 | Journal of Experimental & Clinical Cancer Research

Fig. 3

From: Flavagline analog FL3 induces cell cycle arrest in urothelial carcinoma cell of the bladder by inhibiting the Akt/PHB interaction to activate the GADD45α pathway

Fig. 3

FL3-induced cell cycle inhibition in the G2/M phase via the GADD45α pathway. a After incubation with FL3 for 24 h, T24 cells were harvested and subjected to flow cytometry analysis. The percentage of cells in each phase was presented in the histograms, data represent mean ± SD of three independent experiments, *P < 0.05, **P < 0.01 means significance. b Total cell lysates were isolated from T24 cells treated with concentrations of FL3 for 24 h, and subjected to Western blot analysis with indicated antibodies (left panel). The Western blots have been quantified by densitometry and the quantitative values have been incorporated below the Western blot bands. c Cell cycle distribution of PHB knockdown T24 cells was analyzed by flow cytometry, and the percentage of cells in each phase was shown as histograms, data represent mean ± SD of three independent experiments, *P < 0.05 denotes a significant difference. d Total cell lysates from PHB knockdown or control T24 cells were isolated and subjected to Western blot analysis with the indicated antibodies (left panel). The Western blots have been quantified by densitometry and the quantitative values have been incorporated below the Western blot bands. e Regulated genes after FL3 treatment or PHB knockdown in T24 cells were compared with microarray analysis of mRNA expression. f Commonly regulated genes were analyzed in both microarray analyses, and GADD45α was significantly upregulated in both analyses. g Total cell lysates from FL3-treated or PHB knockdown T24 cells were subjected to Western blot analysis with primary GADD45α antibody, indicating that the protein levels of GADD45α were upregulated. h Quantitative real-time PCR (qRT-PCR) was performed to detect the mRNA expression of GADD45α. *P < 0.05 denotes significant difference compared to control

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