Journal: Nature Communications

Article Title: DNA replication fork speed acts as a pacer in cortical neurogenesis

doi: 10.1038/s41467-025-65269-y

Figure Lengend Snippet: a Schematic map showing MCMBP-mediated assembly, and export MCM3-7 into nuclear which forms nascent MCMs with MCM2 as pre-replication complexes, and regulates DNA replication fork speed. NLS means nuclear localization signal. b Western blot analysis showing the expression of MCMBP, Cyclin E1, and Ki67 during cortical development. Note that the experiment was performed once for each sample. c Western blot analysis showing downregulation of MCMBP expression in the E15.5, E16.5 and P4 cortex. (mea n ± SD, two-tailed unpaired t-test, ctrl: n = 3, cKO: n = 3). d Representative images of cKO mouse and littermate control at P3. The red star indicates cKO mouse. e Analysis of Mendelian ratios from a Mcmbp fl/+ ; Emx1 Cre/+ × Mcmbp fl/fl cross, analyzing 76 mice at E15.5 and 98 mice at P21 (χ2-Test). f (Left) Dorsal view of Mcmbp +/+ ; Emx1 Cre and Mcmbp fl/fl ; Emx1 Cre (cKO) brains at P4. (Right) Quantification of cortical length, width, area and olfactory bulb. (mean, two-tailed unpaired t -test; ctrl: n = 7, cKO: n = 5). g DAPI staining coronal sections and cortical plate thickness analysis of Mcmbp +/+ and cKO P4 brain (mean, two-tailed unpaired t-test, ctrl: n = 7, cKO: n = 5). h Immunostaining of layer markers BRN2, TBR1, LHX2 and TLE4 in P4 brain of Mcmbp +/+ ; Emx1 Cre and cKO. (mean, two-tailed unpaired t-test, BRN2, TBR1, ctrl: n = 8, cKO: n = 5, LHX2, TLE4, ctrl: n = 4, cKO: n = 5). i Immunostaining for layer markers BRN2 and TBR1 in P21 brains. (mean, two-tailed unpaired t -test, ctrl: n = 4, cKO: n = 3). j Immunostaining of apical progenitor marker SOX2 and intermediate progenitor marker EOMES from E12.5 to E16.5. k SOX2+ cell number analysis from E12.5 to E16.5. (mean, two-tailed unpaired t-test, E12.5, ctrl: n = 5, cKO: n = 4, E13.5, ctrl: n = 4, cKO: n = 3, E14.5, ctrl: n = 5, cKO: n = 5, E15.5, ctrl: n = 6, cKO: n = 4, E16.5, ctrl: n = 6, cKO: n = 4). l EOMES+ cell number analysis from E12.5 to E16.5. (mean, two-tailed unpaired t-test, E12.5, ctrl: n = 3, cKO: n = 3, E13.5, ctrl: n = 4, cKO: n = 4, E14.5, ctrl: n = 4, cKO: n = 4, E15.5, ctrl: n = 4, cKO: n = 3, E16.5, ctrl: n = 4, cKO: n = 3).Source data are provided as a Source Data file.

Article Snippet: The following primary antibodies were used for immunoblotting: MCMBP (1:2000, Invitrogen, PA5-58410), MCM2 (1:1000, Proteintech, 66009-1-Ig), MCM3 (1:1000, Santa Cruz Biotechnology, SC-390480), MCM4 (1:1000, Proteintech, 13043-1-AP), MCM5 (1:1000, Proteintech, 11703-1-AP), MCM6 (1:1000, Santa Cruz Biotechnology, SC-393618), MCM7 (1:1000, Proteintech, 11225-1-AP), ARL13B (1:1000, Proteintech, 17711-1-AP), CEP83(1:1000, Proteintech, 26013-1-AP), PCNT (1:2000, Abcam, ab220784), NINEIN (1:1000, Santa Cruz Biotechnology, SC376420), EB3 (1:1000, 1:1000, Proteintech, 23974-1-AP), β-actin (1:2000, Proteintech, 66009-1-Ig), PCNA (1:1000, Servicebio, GB12010), Rabbit Monoclonal anti-Phospho-Chk1 (Ser345) (1:1000, Cell Signaling Technology, 2348), Rabbit Polyclonal anti-Phospho-p53 (Ser15) (1:1000, Invitrogen, PA5-104742), Mouse Monoclonal anti-Chk1 (1:1000, Cell Signaling Technology, 12302), and Rabbit Polyclonal anti-ATR (1:1000, Cell Signaling Technology, 13934), Rabbit Monoclonal anti-Ki67 (1:1000, Abcam, ab16667), Rabbit Polyclonal anti-Cyclin E1 (1:1000, Proteintech, 11554-1-AP) Rabbit Monoclonal anti-P53 (1C12) (1:1000, Cell Signaling Technology, 2524S).

Techniques: Western Blot, Expressing, Two Tailed Test, Control, Staining, Immunostaining, Marker