Fig. 4

Interaction between vitamin C-induced increase in 5-hydroxymethylation and vitamin C-induced loss of H3K9me2. a Western blot for H3K9me2 in wild-type and Tet1/2 double knockout (Tet DKO) ES cells ± vitamin C. Vitamin C-induced loss of H3K9me2 still occurs in Tet DKO ES cells that do not undergo vitamin C-induced increase in 5-hmC. b Dot blot for 5-hydroxymethylcytosine (5-hmC) in wild-type parental TT2, G9a knockout, and GLP knockout ES cells ± vitamin C. Vitamin C-induced increase in 5-hmC still occurs in G9a and GLP knockout ES cells depleted of H3K9me2. c meDIP-qPCR on ES cells ± vitamin C that were untreated, treated with a non-targeting (NT) siRNA, or treated with siRNAs against Kdm3a and Kdm3b. Vitamin C-induced DNA demethylation occurs in the absence of Kdm3a/b and loss of H3K9me2. Data are mean ± SD. Asterisks represent P < 0.05 by t test