PMC

Characterization of the enzymatic properties of free EreB and EreB@modified palygorskite. Effects of: (a) temperature and (b) pH on the relative activity at their own optimal pH (8.0) and temperature (45 °C), respectively. (c) Kinetic analysis of free EreB and immobilized EreB. (d) Recycling performance of EreB@modified palygorskite. The letters above different columns addressed not by the same letter are significantly different according to student’s t test, (p < 0.01).

Preparation of purified EreB: (a) agarose gel electrophoresis: Lane 1: empty plasmid of E. coli; Lane 2: recombinant E. coli; (b) SDS-PAGE electrophoresis: Lane 1: supernatant of the EreB-expressing cells; Lane 2: sediment of the EreB-expressing cells; Lane 3: empty plasmid of E. coli; (c) Lane 1: supernatant of the EreB-expressing cells; Lane 2: purified enzyme; Lane 3: empty plasmid of E. coli.

Restriction maps of the Madrid E gltA locus (top) and predicted pMW1047 insertion in RPMOB.001 (bottom). H, HindIII; P, PstI; X, XbaI. Numbers below the relevant restriction sites indicate the location of the restriction site on the map. Arrows identify the location and orientation of the gltA and ereB genes.

TET2 expression depends on EBNA2 and RBP-jκ in type III latency. (A) EREB cells withdrawn from estradiol (E2) for 24, 48, or 72 h were assayed by Western blotting for TET2, EBNA2, or actin. (B) RT-qPCR results for TET2 mRNA for EREB cells treated as described for panel A. (C) ChIP-qPCR results for EBNA2 at EBNA2_A, EBNA2_B, RBP_L, or RBP_R sites at TET2 loci or at control actin in EREB cells in the presence (+E2) or absence (−E2) of estradiol for 48 h. (D) ChIP-qPCR results for RBP-jκ at RBP_L or RBP_R sites or at control actin in EREB cells in the presence (+E2) or absence (−E2) of estradiol for 48 h. (E) LCLs transduced with three different shRNAs for RBP-jκ or control (shCtrl) and assayed by Western blotting with antibody to TET2, RBP-jκ, or actin. (F) Cells were treated as described for panel E and then assayed by RT-qPCR for TET2 mRNA relative to actin. *, P < 0.05.