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. 2013 May 20;288(27):19882–19899. doi: 10.1074/jbc.M113.473181

FIGURE 8.

FIGURE 8.

H3.3 regulates transcription at the CMV promoter-regulated transgene array. A, strand-specific qRT-PCR analysis of total RNA collected from U2OS 2-6-3/YFP-MS2/rtTA cells expressing full-length FLAG-tagged H3.3 constructs 0 and 3 h after activation with doxycycline (Dox) using a primer pair in rabbit β-globin exon 3. Results are the average of at least three independent experiments. S.D. values, in the form of error bars, and p values, calculated using unpaired t test, are presented in the graphs. Western blot analysis of FLAG-tagged H3.3 constructs using the FLAG antibody and γ-tubulin as a loading control. B, strand-specific qRT-PCR analysis of total RNA collected from U2OS 2-6-3/YFP-MS2/rtTA, expressing control pLKO.1 and H3.3 shRNA constructs, 0 and 3 h after activation with doxycycline. C, qRT-PCR analysis of hH3F3A and hH3F3B mRNA levels in U2OS 2-6-3/YFP-MS2/rtTA cells after knockdown with gene-specific shRNAs. Shown is Western blot analysis of H3.3 levels in control and knockdown cells using an H3.3-specific antibody and γ-tubulin as a loading control.