Additional file 11: of Loss of CREST leads to neuroinflammatory responses and ALS-like motor defects in mice

Figure S6. Validation of Knockdown Efficiency of shRNAs and Inhibitory Effect of CREST on Transcription of Cxcl10 in Cultured Neurons, related to Fig. 5. (A) Immunoblot of protein samples from primary cortical neurons isolated from embryotic Crest −/− and WT littermates, using anti-CREST (top) and anti-Actin (bottom) antibodies. (B) Immunoblot and intensity quantification of protein from primary cortical neurons infected with lentivirus carrying CREST shRNA (shCREST) and GFP (as control). Relative expression represents the intensity ratios of CREST/GAPDH. (C) Immunoblot of protein from primary cortical neurons infected with lentivirus expressing GFP or HA-tagged WT CREST, using anti-HA (top) and anti-GAPDH (bottom) antibodies. (D) Quantitative RT-PCR of Cxcl10 (left) and Ccl2 (right) in primary cortical neurons infected with lentivirus expressing GFP or HA-tagged WT CREST. (E and F) Immunoblot (E) and quantification (F) of protein from primary cortical neurons infected with lentivirus carrying BRG1 shRNA (shBRG1) and GFP (as control). Relative expression represents the intensity ratios of BRG1/Actin normalized to one control sample. Error bars represent SEM. *p < 0.05, ***p < 0.001, and ****p < 0.0001, Student’s t test. (TIF 8284 kb)