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Fig. 5 | BMC Cancer

Fig. 5

From: Antagonists of the serotonin receptor 5A target human breast tumor initiating cells

Fig. 5

Inducible knockout of HTR5A affects tumorsphere formation and targets BTIC. a FACS scatter plot showing the frequency of Cas9-GFP+ tumor cells in an MCF-7 iCas9 cell line grown in dox-containing or dox-free media for 48 h. b Percentage INDEL frequency at each sgRNA-binding locus of HTR5A was obtained by next-generation sequencing of genomic DNA isolated from 3 MCF-7 HTR5A iKO clones grown in dox-containing media or the vehicle for 48 h. c The number of tumorspheres formed in media containing the vehicle (black bars) or dox (white bars) for each cell line was determined over 3 successive passages. Bar graphs display the mean number of tumorspheres +/− the SEM. d Ex vivo assays demonstrate that induction of Cas9 expression in the HTR5A iKO 2–8 cell line (solid lines) delays the onset of tumor xenografts (compare uninduced (black) with dox-induced (green)), whereas induction of Cas9 expression has no effect on the appearance of tumor xenografts in the NT iKO cell line (dotted lines). Statistical significance was determined using Kaplan-Meier survival analysis and the log-rank test [(I) * p = 0.0047; (II) * p = 0.01]

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