Figure S2.
EVL is the predominant Ena/VASP paralog regulating dendritic filopodia. (A) Uniform manifold approximation and projection (UMAP) subcluster categories of single-cell RNA-seq expression profiles from mouse glutamatergic neurons. 1: L6b cortex (CTX), 2: L6 corticothalamic CTX, 3: L5 near-projecting CTX, 4: L6 intratelencephalic (IT) CTX, 5: L5 IT CTX, 6: L5 pyramidal tract CTX, 7: L2/3 IT CTX-1, 8: Car3, 9: L4/5 IT CTX, 10: L2/3 IT CTX Ppp1r18, 11: L2/3 IT CTX-2. (B) Log2(TPM(transcripts per million)) expression in mouse of Enah, Evl, and Vasp across single-cell RNA-seq UMAP subcluster categories. (C) UMAP subcluster categories of single-cell RNAseq expression profiles from human glutamatergic neurons. 1: L5/6 near-projecting cortex (CTX), 2: L6 corticothalamic CTX, 3: intratelencephalic (IT) CTX, 4: L5/6 IT CTX Car3, 5: L6b CTX, 6: L4 IT CTX, 7: L5 IT CTX. (D) Log2(TPM) expression in human of ENAH, EVL, and VASP across single-cell RNA-seq UMAP subcluster categories. (E and F) Comparison of Enah, Evl, and Vasp expression in mouse (E) and human (F) across single-cell RNA-seq grouped by glutamatergic neurons versus non-neuronal cell types. Statistical comparisons of RNA abundance in cell type categories made by Welch’s t test, n = 250 cells. (G) Representative western blot confirming specificity of VASP antibody in NIH3T3 (left) and primary neuron cultures (right) with pLKO-shRNA-TurboRFP targeting Vasp mRNA. Mean ± SD. N = 3 biological replicates. (H and I) Representative western blots of protein lysates from primary neurons at D11, transduced on D7 with indicated pLKO-shRNA-TurboRFP lentiviral particles targeting Enah (H; left) or Evl mRNA (I; left). Fold change quantification of knockdown compared to non-targeting shRNA control (right). Mean ± SD. N = 3 biological replicates. Statistical comparisons of shRNA knockdown efficiency were made by unpaired t test. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, n.s. is not significant. See also Fig. 1 and Video 1. Source data are available for this figure: SourceData FS2.

EVL is the predominant Ena/VASP paralog regulating dendritic filopodia. (A) Uniform manifold approximation and projection (UMAP) subcluster categories of single-cell RNA-seq expression profiles from mouse glutamatergic neurons. 1: L6b cortex (CTX), 2: L6 corticothalamic CTX, 3: L5 near-projecting CTX, 4: L6 intratelencephalic (IT) CTX, 5: L5 IT CTX, 6: L5 pyramidal tract CTX, 7: L2/3 IT CTX-1, 8: Car3, 9: L4/5 IT CTX, 10: L2/3 IT CTX Ppp1r18, 11: L2/3 IT CTX-2. (B) Log2(TPM(transcripts per million)) expression in mouse of Enah, Evl, and Vasp across single-cell RNA-seq UMAP subcluster categories. (C) UMAP subcluster categories of single-cell RNAseq expression profiles from human glutamatergic neurons. 1: L5/6 near-projecting cortex (CTX), 2: L6 corticothalamic CTX, 3: intratelencephalic (IT) CTX, 4: L5/6 IT CTX Car3, 5: L6b CTX, 6: L4 IT CTX, 7: L5 IT CTX. (D) Log2(TPM) expression in human of ENAH, EVL, and VASP across single-cell RNA-seq UMAP subcluster categories. (E and F) Comparison of Enah, Evl, and Vasp expression in mouse (E) and human (F) across single-cell RNA-seq grouped by glutamatergic neurons versus non-neuronal cell types. Statistical comparisons of RNA abundance in cell type categories made by Welch’s t test, n = 250 cells. (G) Representative western blot confirming specificity of VASP antibody in NIH3T3 (left) and primary neuron cultures (right) with pLKO-shRNA-TurboRFP targeting Vasp mRNA. Mean ± SD. N = 3 biological replicates. (H and I) Representative western blots of protein lysates from primary neurons at D11, transduced on D7 with indicated pLKO-shRNA-TurboRFP lentiviral particles targeting Enah (H; left) or Evl mRNA (I; left). Fold change quantification of knockdown compared to non-targeting shRNA control (right). Mean ± SD. N = 3 biological replicates. Statistical comparisons of shRNA knockdown efficiency were made by unpaired t test. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, n.s. is not significant. See also Fig. 1 and Video 1. Source data are available for this figure: SourceData FS2.

This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal