Skip to main content
. 2012 Feb 27;7(2):e32451. doi: 10.1371/journal.pone.0032451

Figure 3. BCR-JAK2 is constitutively phosphorylated, triggering STAT5 activation which translocates to the nucleus and induces STAT5 target gene expression on transduced Ba/F3 cells.

Figure 3

(a) Western blot analysis of Ba/F3 cells infected with BCR-JAK2, TEL-JAK2 or vector alone (mock). Cellular lysates were immunoprecipitated with anti-pTyr Ab and immunoblotted with anti-JAK2 Ab to see pJAK2 (upper panel: endogenous JAK2: 130 kDa, BCR-JAK2: 90 kDa, TEL-JAK2: 72 kDa). TEL-JAK2 protein expression is higher than BCR-JAK2 in transduced Ba/F3 cells. Whole cell lysates were probed with anti-pSTAT5 and anti-STAT5 Ab's (bottom). The expression levels of Tubulin were used as a loading control. (b) Enriched cytoplasmic (C) and nuclear (N) extracts from Ba/F3-mock,Ba/F3-BCR-JAK2 and Ba/F3-TEL-JAK2 cells were prepared from total cell lysates and blotted with anti-pSTAT5 Ab. TATA-Binding protein (TBP) and IkBα were used as loading controls for enriched nuclear and cytoplasmic fractions, respectively. (c) Expression of Bcl-xL, Osm and Socs2 in Ba/F3-mock, Ba/F3-BCR-JAK2 and Ba/F3-TEL-JAK2 cells by qPCR. For comparative purposes, mRNA levels in untreated cells were normalized to 1. Bars represent fold changes of each gene normalized using GADPH levels. Results are given as mean ± SEM (n = 3).