Figure 1.

Polyadenylation of Canonical Histone H3.1 mRNA Enhances Tumor Formation in Nude Mice

(A) An H3.1 cDNA fragment was inserted into the multiple clone sites (MCSs) of pcDNA-FLAG vector, which contains a poly(A) signal in the downstream of MCSs and generates polyadenylated mRNAs.

(B) pcDNA-FLAG-Empty (EV) or FH3.1poly(A) vector was stably transfected into BEAS-2B cells. The amount of total and polyadenylated H3.1 mRNA was measured by RT-qPCR using cDNAs synthesized with random or oligo (dT) primers, respectively. mRNA levels for H3.1 were normalized to Actin. The data shown are the mean ± S.D. (n = 3).

(C) Western blot shows exogenous H3.1 protein generated by transfection of FH3.1poly(A)(left) and the increase in H3 protein following arsenic treatment (right).

(D) Cell growth rate was measured by MTT. The data shown are the mean ± S.D. (n = 3). Student's t test was applied for statistical significance: ∗∗p < 0.01.

(E) Soft agar assays. The data shown are the mean ± S.D. (n = 3). Student's t test was applied for statistical significance: ∗∗p < 0.01.

(F–H) A total of five million transformed or control cell clones were injected into nude mice. After 5 months postinjection, the mice were sacrificed. Tumor number (F), volume (G), and weight (H) were calculated. Tumor diameters were measured with calipers, and the tumor volume was calculated. Results represent the mean ± S.D. (n = 18). Student's t test was applied for statistical significance: ∗p < 0.05.