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Additional Supporting Information may be found in the online version of this article.

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sc-12-0742_sm_SupplData.pdf7KSupplementary Data
sc-12-0742_sm_SupplFigure1.tif229KSupplementary figure 1. Lineage trees reconstructed from single dorsal pallial progenitors from Emc1-cre; ZEG mice. The numbers of recording intervals were indicated in lineage trees. The PN lineage was indicated by red lines and the IN lineage was indicated by green lines. (A1-A2) These two lineage trees showing that PN were determined earlier than IN.
sc-12-0742_sm_SupplFigure2.tif1174KSupplementary figure 2. Lineage trees reconstructed from single pallial progenitors from Dlx5/6-CIE mice. (A1-A6) PN were determined earlier than IN. (B1-B3) PN were determined later than IN. (C1-C5) PN and IN were determined at the same time.
sc-12-0742_sm_SupplFigure3.pdf113KSupplementary figure 3. Lineage trees reconstructed from single pallial progenitors from GAD67-GFP mice. (A1-A9) PN were determined earlier than IN. (B1-B3) PN were determined later than IN. (C1-C4) PN and IN were determined at the same time. Note that the lineage tree in A3 showing that many PN were generated first, and then one IN was generated.
sc-12-0742_sm_SupplFigure4.tif1997KSupplementary figure 4. Individual dorsal pallial progenitors from E13.5 GAD67-GFP mice generated both projection neuron-like and interneuron-like cells. (A1-A8) Sequential phase-contrast images showing that one progenitor (P, white arrow in A1, non-GFP progenitor) developed into a clone. The cells that were not generated from the progenitor were indicated by white arrowheads. (B1-B8) Fluorescence images of A1-A8. Note that green (GFP) signals (green arrows) were observed in B5-B8. (C1-C4) Three Tuj1+/GFP-negative cells (red arrows) and three Tuj1+/GFP+ cells (green arrows) were observed in this clone. Recording time in hrs:mins was indicated in upper right corner of each panel. Scale bars: 100 μm in A1-B8, 50 μm in C1-C4.
sc-12-0742_sm_SupplFigure5.tif1404KSupplementary figure 5. Individual dorsal pallial progenitors from E15.5 GAD67-GFP mice generated both projection neuron-like and interneuron-like cells. (A1-A10) Sequential phase-contrast images showing that one progenitor (P, white arrow in A1, non-GFP progenitor) developed into a clone. The cells that were not generated from the progenitor were indicated by white arrowheads. (B1-B5) Fluorescence images of A6- A10. Note that the green (GFP) signals (green arrows) were observed in B2-B5. (C1-C3) Two Tuj1+/GFP-negative cells (red arrows) were observed in this clone. Green circles were used to indicate the GFP+ cells that were washed off during the immunocytochemistry staining process. Scale bars: 100 μm in A1-B5, 50 μm in C1-C3.
sc-12-0742_sm_SupplFigure6.tif880KSupplementary figure 6. Individual dorsal pallial progenitors from E17.5 GAD67-GFP mice generated both projection neuron-like and interneuron-like cells. (A1-A6) Sequential phase-contrast images showing that one progenitor (P, white arrow in A1, non-GFP progenitor) developed into a clone. The cells that were not generated from this progenitor were indicated by white and green arrowheads. (B1-B6) Fluorescence images of A1-A6. The green (GFP) signals (green arrows) were observed in B2-B6. Note that one green (GFP) cell (green arrowhead in A1, B1) was postmitotic. (C1-C4) One Tuj1+/GFP-negative cell (red arrow) and one Tuj1+/GFP+ cell (green arrow) were observed in this clone. Scale bars: 100 μm in A1-B6, 50 μm in C1-C4.
sc-12-0742_sm_SupplFigure7.tif666KSupplementary figure 7. Individual dorsal pallial progenitors from E17.5 GAD67-GFP mice generated both projection neuron-like and interneuron-like cells. (A1-A8) Sequential phase-contrast images showing that one progenitor (P, white arrow in A1, non-GFP progenitor) developed into a clone. The cell that was not generated from the progenitor was indicated by green arrowhead. (B1-B8) Fluorescence images of A1-A8. The green (GFP) signals (green arrows) were observed in B4-B8. Note that one green (GFP) cell derived from the progenitor divided into two green (GFP) cells (vertical green arrows in A4-6, B4-6). There was a postmitotic green (GFP) cell (green arrowhead in A1, B1) at the beginning of recording. (C1-C4) One Tuj1+/GFP-negative cells (red arrow) and four Tuj1+/GFP+ cells were observed in this clone. The green circle was used to indicate the GFP+ cell that was washed off during the immunocytochemistry staining process. Scale bars: 50 μm.
sc-12-0742_sm_Suppltable1.pdf13KSupplementary Table 1. Number of single pallial progenitor-derived clones from E11.5 mouse brains. From time-lapse videos, we totally identified 112 clones derived from individual pallial progenitors from E11.5 Emx1-cre; Z/EG mice. 47 clones contained GFP+/Sp8+/CR+/Tuj1+ interneuron-like cells (IN). 13 clones contained both GFP+/Tbr1+/Tuj1+ projection neuron-like cells (PN) and IN. We also identified 999 clones derived from individual pallial progenitors from E11 Dlx5/6-CIE or GAD67-GFP mice. About 42.6% of them (426 clones) contained IN (Tuj1+/GFP+ cells); 135 clones contained both PN (non-GFP but Tuj1+) and IN. 40 lineage trees were successfully reconstructed from clones that contained both projection neuron-like and interneuron-like cells (Fig. 6, and supporting information Fig. 1-3); the remainder of the clones were difficult to follow, either due to the close proximity of some progenies (making it impossible to accurately distinguish them; see supplemental online video 2); or some progenies washed off the dish during the immunocytochemistry staining process.
sc-12-0742_sm_Suppltable2.pdf10KSupplementary Table 2. Number of single pallial progenitor-derived clones from E13.5, E15.5 and E17.5 GAD67-GFP mouse brains. From time-lapse videos, we totally identified 842, 496 and 602 clones derived from individual pallial progenitors from E13.5, E15.5, E17.5 GAD67-GFP mice. Only 0.007% of total clones (14/1940) contained both Tuj1+/GFP+ interneuron-like cells (IN) and Tuj1+/GFP-negative projection neuron-like cells (PN).
sc-12-0742_sm_onlinevideo1.mov4563KSupplemental online video 1. This time-lapse video was recorded for 154 hrs and showed that one pallial progenitor from E11.5 Emx1-cre; Z/EG mice developed into a clone in vitro. At the beginning of recording, there were two cells very close to each other. The upper cell was a progenitor cell (P, white arrow) that developed into a clone. This clone contained both projection neuron-like (red arrows) and interneuron-like cells (see Fig. 3). The lower one (arrowhead) was a postmitotic cell.
sc-12-0742_sm_onlinevideo2.mov4193KSupplemental online video 2. This time-lapse video showing that one pallial progenitor from E11.5 Dlx5/6-CIE mice developed into a clone. At the beginning of recording, there were two cells very close to each other. The upper cell was a progenitor cell (P, white arrow, non-GFP progenitor) that developed into a clone. After three rounds of division, weak green (GFP) signals from two progenies were detected (green arrows, t =57:00); one died later (t =136:53). Another progeny (left green arrow) with weak GFP signal appeared approximately 6 days later after plating (t =146:53). This clone contained both projection neuron-like and interneuron-like cells (see Fig. 4). Arrowheads in the left video indicated the cells that were not derived from this progenitor cell.
sc-12-0742_sm_onlinevideo3.mov4129KSupplemental online video 3. This video showed that one pallial progenitor from E11.5 GAD67-GFP mouse forebrains developed into a clone. This clone contained 2 projection neuron-like and 3 interneuron-like cells (see Fig. 5). The green (GFP) signals from 3 GFP+ cells (green arrows) were detected soon after they became postmitotic (t =43:35).
sc-12-0742_sm_onlinevideo4.mov4367KSupplemental online video 4. This video showed that one dorsal pallial progenitor from E13.5 GAD67-GFP mouse forebrains developed into a clone. At the beginning of recording, there were two cells very close to each other. The upper cell was a progenitor cell (P, white arrow, non-GFP progenitor) that developed into a clone. Arrowheads in the left video indicated the cells that were not derived from this progenitor cell. The green (GFP) signals from 2 GFP+ cells (green arrows) were detected 55 hrs after recording. This clone contained 3 projection neuron-like cells and 3 interneuron-like cells (see supporting information Fig. 4).
sc-12-0742_sm_onlinevideo5.mov4095KSupplemental online video 5. This video showed that one dorsal pallial progenitor from E15.5 GAD67-GFP mouse forebrains developed into a clone. The progenitor cell (P, white arrow, non-GFP progenitor) developed into a clone, which contained 2 projection neuron-like cells and 2 interneuron-like cells (see supporting information Fig. 5). The green (GFP) signals from 2 GFP+ cells (green arrows) were detected 50 hrs after recording. Two postmitotic cells (arrowheads) also existed at the beginning of recording.
sc-12-0742_sm_onlinevideo6.mov4092KSupplemental online video 6. This video showed that one dorsal pallial progenitor from E17.5 GAD67-GFP mouse forebrains developed into a clone. The progenitor cell (P, white arrow, non-GFP progenitor) developed into 1 projection neuron-like cell (white arrow) and 1 interneuron-like cell (green arrow) (see supporting information Fig. 6). The green (GFP) signal from the GFP+ cell was detected soon after it became postmitotic (t =20:20). One postmitotic green cell (green arrowhead) existed at the beginning of recording.
sc-12-0742_sm_onlinevideo7.mov4117KSupplemental online video 7. This video showed that one dorsal pallial progenitor from E17.5 GAD67-GFP mouse forebrains developed into a clone. The progenitor cell (P, white arrow, non-GFP progenitor) developed into a clone that contained 1 projection neuron-like cell and 5 interneuron-like cells (see supporting information Fig. 7). The green (GFP) signals from the GFP+ cells (green arrow) were detected 65 hrs after recording. Note that one green (GFP) cell derived from this progenitor divided into two green (GFP) cells (vertical green arrows). Two postmitotic cells (green and white arrowhead) existed at the beginning of recording.

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