(A) qPCR analysis of Khdrbs1 mRNA levels in the cortex of embryonic (E10.5-E15.5) and post-natal (P0-25dpp) mouse brain. Khdrbs1 relative expression was evaluated by △CT method using L34 expression …
(A) Schematic representation of the stratification of the VZ/SVZ, intermediate zone (IZ) and cortical plate (CP) in the E13.5 embryonic cortex. Molecular markers, expressed by the respective cell …
(A) Representative images of immunofluorescence analysis of BrdU (left panel) and SOX2(center panel) performed in E13.5 cortex of embryos treated with BrdU for 2 hr. Bar graphs represent (mean±SD) …
(A) Diagram of the experimental design to investigate the fate on NPCs during neurogenesis. Pregnant females were treated with BrdU at E12.5, and embryos were collected after 2 days for …
Weight analysis of Khdrbs1+/+, Khdrbs1+/- and Khdrbs1-/- mouse embryos at E13.5, 15.5 and E17.5. At least 5 embryos for each genotype were weighted. Statistical analysis was performed using the …
(A) Bright field images of Khdrbs1+/+ and Khdrbs1-/- NPCs cultured in proliferating condition for 3 days in culture. The insets show higher magnification of a selected neurosphere. Black arrowheads …
List of the genes upregulated and downregulated inSam68 knockout NPCs.
Yellow=upregulated genes; green=downregulated genes. NPCs, Neural progenitor cells.
List of exons regulated by SAM68 in mouse NPCs.
Green: genes downregulated at expression level; Yellow: genes up-regulated at expression level; White: genes unchanged at expression level. NPCs, Neural progenitor cells.
(A) Bar graph reporting the analysis of the diameter of neurospheres formed by Khdrbs1+/+ and Khdrbs1-/- NPCs in 1–3 days of culture. N = 3; *p<0.05. (B) Cell death analysis of Khdrbs1+/+ and Khdrbs1…
(A) Images representing a typical neuron obtained through the differentiation of Khdrbs1-/- NPCs (upper image) and the representation of the scheme used to quantify the complexity of its structure …
(A) Analysis of the gene and exon expression in Khdrbs1-/- vs Khdrbs1+/+ NPCs reporting upregulated and downregulated events. (B) Overlap between the genes regulated at expression and splicing level …
(A) Diagram of ALDH1A3 gene structure. Red and green squares represent exons, respectively, upregulated and downregulated in the microarray analysis. Proximal polyadenylation site (pPAS) and distal …
(A) Diagramatic scheme of the Aldh1a3 gene reporting the up- (red) and down-regulated (green) exons identified by the microarray experiment. The bar graphs represent the results of qPCR analyses …
(A) Western blot analysis of the expression of ALDH1A3 and Sam68 in lysates from Khdrbs1+/+ and khdrbs1-/- NPCs. Vinculin was used as loading control. The bar graph represent densitometric analyses …
(A) Western blot analysis of ALDH1A3△ expression in Khdrbs1-/- NPCs treated with 10 µM proteasome inhibitor MG132 and 2 µM autophagy inhibitor Chloroquine. HEK293 cells transfected with pCI-ALDH1A3△ …
(A) Representative images of Khdrbs1+/+ NPCs transfected with GFP vector and Khdrbs1-/- NPCs transfected with GFP and GFP-ALDH1A3 vectors, cultured for 3 days in proliferating condition. Scale bar …
(A) Representative images of Khdrbs1-/- NPCs transfected with GFP vector and Khdrbs1+/+ NPCs transfected with GFP and GFP-ALDH1A3△ vectors, cultured for 3 days in proliferating condition. Scale bar …
(A) Evaluation of the relative lactate production of Khdrbs1+/+ and Khdrbs1-/- NPCs cultured under proliferating condition. (B) Relative lactate production in Khdrbs1+/+ or Khdrbs1-/- NPCs treated …
Schematic model of the function of Sam68 in the modulation of NPCs stemness. High Sam68 expression in NPCs of the neocortex insures high expression of ALDH1A3 protein and anaerobic glycolysis, thus …
List of the oligonucleotides used as PCR primers and siRNAs in the study.