Fig. 2

ZDHHC15 isoform expression in GBM cells and GSCs. a Diagram of the ZDHHC15 splicing isoforms. The isoform 1 coding sequence contains 12 exons. Compared to isoform 1, variant 2 lacks coding for exon 2, and variant 3 lacks an in-frame coding exon and differs at the 3′ end. The locations of the primers and the sites targeted with stealth siRNAs are indicated on a diagram of the ZDHHC15 splicing isoforms. b RT-PCR analysis of the mRNA levels of ZDHHC15 splicing isoforms in six GBM cells (H4, A172, U87, T98G, U251, and LN18). Sequences encoding isoforms 1 and 2 using primers #1 and #3 (compared to isoform 1, variant 2 lacks 27 bp). Sequences encoding isoform 1 used primers #2 and #3. Sequences encoding isoform 2 used primers #4 and #5. Sequences encoding isoform 3 used primers #6 and #7. β-Actin served as the loading control. c RT-PCR analysis of the mRNA levels of ZDHHC15 splicing isoforms in GSCs and NSCs. β-Actin served as the loading control. d RT-PCR analysis of the mRNA levels of ZDHHC15 splicing isoforms during GSC self-renewal and the differentiation stage. β-Actin served as the loading control. e Western blot analysis of ZDHHC15 in six GBM cell lines (H4, A172, U87, T98G, U251, and LN18); four GSCs derived from U87, T98G, U251, and LN18; and U251 GSCs after transfection with ZDHHC15 stealth siRNAs. β-Actin served as the loading control