Extended Data Fig. 1: Expression of MEF2C in the human brain and design of mouse Mef2c shRNA constructs.
From: Whole-brain in vivo base editing reverses behavioral changes in Mef2c-mutant mice
(a) Annotation of the genomic and transcript information of L35P (based on the hg38 database). (b) Human brain transcriptomic data for MEF2C was obtained from the Human Brain Transcriptome (HBT), which ranges from embryonic development (periods 3-7) to postnatal development (periods 8–12) to adulthood (periods 13–15). Brain region abbreviations as follows, Hippocampus (HIP), neocortex (NCX), cerebellar cortex (CBC), mediodorsal nucleus of the thalamus (MD), striatum (STR), and amygdala (AMY). (c) Western blotting of MEF2C WT, L35F, L35P, L35I, or L35A expressed in HEK293 cells. (d) Quantifying relative MEF2C protein levels normalized to GAPDH (n = 7 per group). WT versus L35F: P < 0.0001; WT versus L35P: P < 0.0001; WT versus L35I: P = 0.0005; WT versus L35A: P = 0.0003, One-way ANOVA. (e) Schematic illustration of two Mef2c shRNA designed based on BLOCK-iT™ RNAi Designer (http://rnaidesigner.thermofisher.com/rnaiexpress/). (f) Quantifying Mef2c mRNA level of cultured primary cortical neurons infected with lentiviral vectors expressing scrambled or two Mef2c shRNA (P < 0.0001, n = 4 per group, One-way ANOVA). (g) Representative western blotting of cultured primary cortical neurons infected with lentiviral vectors expressing scrambled or Mef2c shRNA-1 for 72 h. Cells were harvested at DIV5. GAPDH was used as the internal control. (h) Quantitative analysis of relative Mef2c protein expression. Neurons infected with scrambled shRNA were normalized to 100% (P = 0.0327, n = 3, unpaired two-tailed Student’s t-test). n is biological repeat numbers of independent experiments. Statistical values represent the mean ± s.e.m. *P < 0.05, ***P < 0.001, ****P < 0.0001.
