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. Author manuscript; available in PMC: 2018 Dec 1.
Published in final edited form as: Mol Cancer Ther. 2017 Sep 27;16(12):2862–2870. doi: 10.1158/1535-7163.MCT-17-0130

Figure 3.

Figure 3

PSMC6 was validated as a BTZ-sensitive gene. (A) A pair of sgRNAs targeting intron region flanking Exon 1. (B) PSMC6 exon 1 deletion was validated by PCR for two cloned cells. (C) PSMC6 expression levels detected by quantitative PCR. (D) Sanger sequencing analyze cloned RPMI8226 cells deleted PSMC6. PCR amplified PSMC6 exon 1 region and cloned into T-vector. Then purified plasmid was analyzed by Sanger sequencing. (E) MTT assay for cloned RPMI8226 cells (clone A and B) deleted PSMC6. The cells were treated with BTZ for 72 hours. Control=RPMI8226 cells infected with lenti-vector with non-specific sgRNA. (F) Over expression of PSMC6 with HA tag in cloned PSMC6 deficiency cells. (G) Over expression of PSMC6 wild type or with HA tag rescues the phenotype of PSMC6 deficiency. NS= non-specific sgRNA control. bortezomib (BTZ). Vector=empty lentivirus vector. PSMC6HA=lentivirus expressing PSMC6 with HA tag.