Figure 1

Shotgun proteomics screen identifies proteins in proliferating E12.5 NSCs including HMGB chromatin proteins. (A-D) Neurospheres containing proliferating E12.5 NSCs express stem cell markers. (E-F) NSCs are multipotent and differentiate into βIII tubulin+ neurons, GFAP+ astrocytes, and CNP+ oligodendrocytes. (G) Heat map of protein expression of 20 prominent proteins in proliferating E12.5 NSCs following passage 2 (P2) and passage 8 (P8) with high (red, >30 spectral counts), medium (yellow, 10-30 spectral counts) and low (white, 1-10 counts) levels of expression. #1 and #2 spectra refer to biological replicates, and the heat map is split into membrane fraction (M), total cell lysate (T) and soluble/supernatant (S). (H) Immunoblot validation of E12.5 proliferating NSC for proteins identified by the proteomics screen, including Matrin3, TAR DNA binding protein (TARDBP/TDP43), HP1 gamma, and HMGB1 and B2. 30 μg of total protein were loaded on the gel. (I) RT-PCR analysis of HMGB1, 2, 3, and 4 mRNA expressions in proliferating E12.5 NSCs. (J) Cellular spatial classification of the proteins identified by the screen.