Fig. 1.

Bcl2L12 up-regulates αB-crystallin protein levels in vitro and in vivo. (A) Western blot analysis of postmitochondrial apoptosis effectors in Ink4a/Arf^−/− astrocytes ectopically expressing a pBabe control, Bcl2L12^V5, LacZ^V5, and Bcl-2. The migration positions of Hsp70, Hsp90, αB-crystallin, Hsp27, and XIAP are indicated. (B) Quantification of αB-crystallin positivity in U98MG xenograft tumor sections. A total of 10–15 low power fields (LPFs) per genotype were counted. Error bars represent SDs, and P values were calculated by using Student's t test. (C) Coexpression analysis of αB-crystallin and Bcl2L12 in primary human GBM. GBM cores were stained with a monoclonal anti-αB-crystallin antibody (red) and a polyclonal Bcl2L12 (anti-L12–2) antiserum (brown). Shown are three representative cores at ×40 magnifications. (D) Heat map (Top; yellow dots = Bcl2L12 positive; red dots = αB-crystallin positive) and density plot analyses (Middle) of Bcl2L12/αB-crystallin double stainings in three representative GBM cores shown in C. The percentages of Bcl2L12 and αB-crystallin-positive and -negative tumor cells are indicated (Bottom) with Bcl2L12 intensities plotted on the x axis and αB-crystallin intensities on the y axis. (E) Analysis of coexpression across 61 GBM cores in comparison with normal brain. Shown are intensity levels of Bcl2L12 (yellow), αB-crystallin (red), and levels of coexpression (green). Pearson correlation coefficient ρ = 0.62.