Figure 1 - supplement 1

Styxl2 was downstream target of Jak1-Stat1 pathway. (A) Multiple sequence alignment of the region surrounding the active motif of the dual-specificity phosphatase catalytic (DSPc) domain among mouse dual-specificity phosphatases and Styxl2. The conserved Cys in functional phosphatases was highlighted in yellow. (B, C) C2C12 cells were transfected with various siRNAs as indicated. After growing in growth medium (GM) for 24 hr (h), cells were harvested either right away (GM 24 h) or after growing in differentiation medium (DM) for an additional 12 hr (DM 12 h). The total RNA and soluble whole cell lysates were subjected to RT-qPCR (B) or Western blot analysis (C), respectively. (D, E) C2C12 cells were treated with 10 ng/ml of leukemia inhibitory factor (LIF) or oncostatin M (OSM) for various times. Cell extracts were subjected to Western blot analysis. (F) Mouse embryos at E9.5 and E10.5 were subjected to in situ hybridization using either sense (control) or antisense probe specific for mouse Styxl2. (G) C2C12 cells were transfected with constructs expressing Flag- or HA-tagged Styxl2 for 24 hr. Cells were then fixed and subjected to immunostaining for Flag or HA. (H) Zebrafish zygotes were co-injected with a plasmid encoding GFP fused with the sequence targeted by Styxl2-morpholino oligo (MO) at the start codon together with or without Styxl2-MO. The phase-contrast and fluorescent images were taken and the representative images were shown