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. 2020 May 6;20(1):533–540. doi: 10.3892/ol.2020.11587

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Copyright: © Bobrowicz et al.

This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.

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Figure 3. — HDAC6 knock-out sensitizes the HUT78 CTCL cell line to PI3K inhibitors. The CTCL cell line, HUT78, was stably transduced with LentiCRISPRv.2 plasmid encoding sgRNA targeting HDAC6 (sequence A and B). sgCON was used as a non-targeting control. (A) Whole-cell lysates from pooled transduced cells were assessed for HDAC6 and acetylated tubulin (a hallmark of HDAC6 inhibition) by western blot analysis. β-actin was used as a loading control. (B) Pooled transduced cells were incubated for 48 h with the investigated PI3K inhibitors and their viability and proliferation were assessed with Cell Titer Glo^®. (C) Whole cell lysates from the cells that underwent selection with puromycin (clones sg9 and sg10) were assessed for HDAC6 and acetylated tubulin (a hallmark of HDAC6 inhibition) by western blot analysis. β-actin was used as a loading control. (D) Cells were incubated for 48 h with the investigated PI3K inhibitors and their viability and proliferation assessed with Cell Titer Glo^®. Statistical significance was assessed by two-way ANOVA with a Bonferroni post-hoc test. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001 vs. sgCON. CTCL, cutaneous T-cell lymphoma; HDAC6, histone deacetylase 6; sgCON, Sg sequence targeting GFP.

Figure 3. — HDAC6 knock-out sensitizes the HUT78 CTCL cell line to PI3K inhibitors. The CTCL cell line, HUT78, was stably transduced with LentiCRISPRv.2 plasmid encoding sgRNA targeting HDAC6 (sequence A and B). sgCON was used as a non-targeting control. (A) Whole-cell lysates from pooled transduced cells were assessed for HDAC6 and acetylated tubulin (a hallmark of HDAC6 inhibition) by western blot analysis. β-actin was used as a loading control. (B) Pooled transduced cells were incubated for 48 h with the investigated PI3K inhibitors and their viability and proliferation were assessed with Cell Titer Glo^®. (C) Whole cell lysates from the cells that underwent selection with puromycin (clones sg9 and sg10) were assessed for HDAC6 and acetylated tubulin (a hallmark of HDAC6 inhibition) by western blot analysis. β-actin was used as a loading control. (D) Cells were incubated for 48 h with the investigated PI3K inhibitors and their viability and proliferation assessed with Cell Titer Glo^®. Statistical significance was assessed by two-way ANOVA with a Bonferroni post-hoc test. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001 vs. sgCON. CTCL, cutaneous T-cell lymphoma; HDAC6, histone deacetylase 6; sgCON, Sg sequence targeting GFP.