PMC:6085830 / 12220-13364 JSONTXT

{"target":"http://pubannotation.org/docs/sourcedb/PMC/sourceid/6085830","sourcedb":"PMC","sourceid":"6085830","source_url":"https://www.ncbi.nlm.nih.gov/pmc/6085830","text":"FIGURE 2: Custom anti-FUS(pSer26) and anti-FUS(pSer30) antibodies are specific to phosphorylated FUS. (A, B) Lysates of HEK293T cells treated with various reagents (DMSO, 50 nM calicheamicin, 100 nM calyculin-A, 200 μM etoposide, 10 μM camptothecin, 2 μM doxorubicin, or 100 nM okadaic acid) for 3 h at 37°C were analyzed by Western blotting with commercial FUS or with anti-FUS(pSer26) or anti-FUS(pSer30) antibodies, respectively; n = 2. (C) Phosphorylated and unphosphorylated synthetic peptides—corresponding to regions within FUS’s prionlike domain—were serially diluted, spotted on nitrocellulose, and immunoprobed with custom antibodies: anti-FUS(pSer26) or anti-FUS(pSer30); n = 2. (D) FUS was knocked down using siRNA in H4 cells then treated with calicheamicin (or DMSO; negative control) to induce phosphorylation. Western blots using anti-FUS(pSer26), anti-FUS(pSer30), and commercial FUS antibodies revealed specificity of the phosphoantibodies to the FUS protein; n = 4. (E) Densitometry analysis of the percentage of signal reduction with FUS knockdown compared with the control; error bars represent 95% confidence intervals. ","tracks":[]}