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. 2022 Oct 27;29(1):271–278. doi: 10.1158/1078-0432.CCR-22-1851

Figure 1.

Figure 1. Potent and specific shRNA-mediated knockdown of DNAJB1-PRKACA. A, Generation of stable Huh7.5 cells expressing the chimeric transcript and shRNAs by serial lentiviral transduction. Sequences of shRNAs tiling over the fusion junction, red and blue letters denote DNAJB1 and PRKACA sequence, respectively. A Ctrl shRNA was used for all experiments. B, Western blot analysis of Huh7-Chimera cells stably transduced with shRNAs A-T, Ctrl shRNA or nontransduced (NT). C, qPCR results of Huh7-Chimera cells stably expressing shRNAs L-P, Ctrl shRNA or untreated cells (UTC). Left graph measures Chimera knockdown, middle graph measures PRKACA knockdown, and right graph measures DNAJB1 knockdown. N = 3 for all conditions, one-way ANOVA with Dunnett multiple comparison test. **, P = 0.0021; **, P = 0.0002; ***, P, ****, P <0.0002.

Potent and specific shRNA-mediated knockdown of DNAJB1-PRKACA. A, Generation of stable Huh7.5 cells expressing the chimeric transcript and shRNAs by serial lentiviral transduction. Sequences of shRNAs tiling over the fusion junction, red and blue letters denote DNAJB1 and PRKACA sequence, respectively. A Ctrl shRNA was used for all experiments. B, Western blot analysis of Huh7-Chimera cells stably transduced with shRNAs A-T, Ctrl shRNA or nontransduced (NT). C, qPCR results of Huh7-Chimera cells stably expressing shRNAs L-P, Ctrl shRNA or untreated cells (UTC). Left graph measures Chimera knockdown, middle graph measures PRKACA knockdown, and right graph measures DNAJB1 knockdown. N = 3 for all conditions, one-way ANOVA with Dunnett multiple comparison test. **, P = 0.0021; ***, P = 0.0002; ****, P < 0.0001.