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(A, B) EBs were generated at day 0 after either d4 (A) or d10 (B) NMP formation and then treated with RA for 10 days (control). In the experimental samples, BMPs were added at day 1 (BMP4+1), day 2 (BMP4+2) or day 4 (BMP4+4) after EB formation, until day 10. Samples were further differentiated for 6 days and then processed for immunohistochemistry (IHC). (C) EBs were labeled with antibodies against LHX2 (dI1, green), FOXD3 (dI2, white), <t>ISL1</t> (dI3, red) and DAPI (all nuclei, blue). (D, E) Quantification of EBs formed from either d4 (D) or d10 (E) NMPs, suggests that dI1s form robustly from the BMP4+1 condition for both d4 and d10 NMPs. In contrast, dI3s form most robustly from BMP4+4 condition for d4 NMPs and the BMP4+1 condition for d10 NMPs. Probability of similarity between control and experimental groups: * p<0.05, ** p<0.005, *** p<0.0005; two-way ANOVA.
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(A, B) EBs were generated at day 0 after either d4 (A) or d10 (B) NMP formation and then treated with RA for 10 days (control). In the experimental samples, BMPs were added at day 1 (BMP4+1), day 2 (BMP4+2) or day 4 (BMP4+4) after EB formation, until day 10. Samples were further differentiated for 6 days and then processed for immunohistochemistry (IHC). (C) EBs were labeled with antibodies against LHX2 (dI1, green), <t>FOXD3</t> (dI2, white), ISL1 (dI3, red) and DAPI (all nuclei, blue). (D, E) Quantification of EBs formed from either d4 (D) or d10 (E) NMPs, suggests that dI1s form robustly from the BMP4+1 condition for both d4 and d10 NMPs. In contrast, dI3s form most robustly from BMP4+4 condition for d4 NMPs and the BMP4+1 condition for d10 NMPs. Probability of similarity between control and experimental groups: * p<0.05, ** p<0.005, *** p<0.0005; two-way ANOVA.
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(A, B) EBs were generated at day 0 after either d4 (A) or d10 (B) NMP formation and then treated with RA for 10 days (control). In the experimental samples, BMPs were added at day 1 (BMP4+1), day 2 (BMP4+2) or day 4 (BMP4+4) after EB formation, until day 10. Samples were further differentiated for 6 days and then processed for immunohistochemistry (IHC). (C) EBs were labeled with antibodies against LHX2 (dI1, green), <t>FOXD3</t> (dI2, white), ISL1 (dI3, red) and DAPI (all nuclei, blue). (D, E) Quantification of EBs formed from either d4 (D) or d10 (E) NMPs, suggests that dI1s form robustly from the BMP4+1 condition for both d4 and d10 NMPs. In contrast, dI3s form most robustly from BMP4+4 condition for d4 NMPs and the BMP4+1 condition for d10 NMPs. Probability of similarity between control and experimental groups: * p<0.05, ** p<0.005, *** p<0.0005; two-way ANOVA.
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(A, B) EBs were generated at day 0 after either d4 (A) or d10 (B) NMP formation and then treated with RA for 10 days (control). In the experimental samples, BMPs were added at day 1 (BMP4+1), day 2 (BMP4+2) or day 4 (BMP4+4) after EB formation, until day 10. Samples were further differentiated for 6 days and then processed for immunohistochemistry (IHC). (C) EBs were labeled with antibodies against LHX2 (dI1, green), <t>FOXD3</t> (dI2, white), ISL1 (dI3, red) and DAPI (all nuclei, blue). (D, E) Quantification of EBs formed from either d4 (D) or d10 (E) NMPs, suggests that dI1s form robustly from the BMP4+1 condition for both d4 and d10 NMPs. In contrast, dI3s form most robustly from BMP4+4 condition for d4 NMPs and the BMP4+1 condition for d10 NMPs. Probability of similarity between control and experimental groups: * p<0.05, ** p<0.005, *** p<0.0005; two-way ANOVA.
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(A, B) EBs were generated at day 0 after either d4 (A) or d10 (B) NMP formation and then treated with RA for 10 days (control). In the experimental samples, BMPs were added at day 1 (BMP4+1), day 2 (BMP4+2) or day 4 (BMP4+4) after EB formation, until day 10. Samples were further differentiated for 6 days and then processed for immunohistochemistry (IHC). (C) EBs were labeled with antibodies against LHX2 (dI1, green), FOXD3 (dI2, white), ISL1 (dI3, red) and DAPI (all nuclei, blue). (D, E) Quantification of EBs formed from either d4 (D) or d10 (E) NMPs, suggests that dI1s form robustly from the BMP4+1 condition for both d4 and d10 NMPs. In contrast, dI3s form most robustly from BMP4+4 condition for d4 NMPs and the BMP4+1 condition for d10 NMPs. Probability of similarity between control and experimental groups: * p<0.05, ** p<0.005, *** p<0.0005; two-way ANOVA.

Journal: bioRxiv

Article Title: In vivo human embryonic spinal cord atlas validates stem cell–derived human dorsal interneurons and reveals ASD spinal signatures

doi: 10.64898/2025.12.22.696129

Figure Lengend Snippet: (A, B) EBs were generated at day 0 after either d4 (A) or d10 (B) NMP formation and then treated with RA for 10 days (control). In the experimental samples, BMPs were added at day 1 (BMP4+1), day 2 (BMP4+2) or day 4 (BMP4+4) after EB formation, until day 10. Samples were further differentiated for 6 days and then processed for immunohistochemistry (IHC). (C) EBs were labeled with antibodies against LHX2 (dI1, green), FOXD3 (dI2, white), ISL1 (dI3, red) and DAPI (all nuclei, blue). (D, E) Quantification of EBs formed from either d4 (D) or d10 (E) NMPs, suggests that dI1s form robustly from the BMP4+1 condition for both d4 and d10 NMPs. In contrast, dI3s form most robustly from BMP4+4 condition for d4 NMPs and the BMP4+1 condition for d10 NMPs. Probability of similarity between control and experimental groups: * p<0.05, ** p<0.005, *** p<0.0005; two-way ANOVA.

Article Snippet: The following antibodies were used: LHX2 (mouse; DSHB, PCRP-LHX2-1C11; 1:50), FOXD3 (guinea pig, a gift from Thomas Mueller, Germany, 1:10,000), ISL1 (goat; R&D systems, AF1837; 1:500), PAX2 (rabbit; Invitrogen, 71-6000; 1:500), LMX1B (guinea pig; Thomas Muller Lab), LHX1/5 (mouse; DSHB, 4F2; 1:50) for 12–14 hours (overnight) at 2–8°C in a humidified chamber.

Techniques: Generated, Control, Immunohistochemistry, Labeling

(A, B) EBs were generated at day 0 after either d4 (A) or d10 (B) NMP formation and then treated with RA for 10 days (control). In the experimental samples, BMPs were added at day 1 (BMP4+1), day 2 (BMP4+2) or day 4 (BMP4+4) after EB formation, until day 10. Samples were further differentiated for 6 days and then processed for immunohistochemistry (IHC). (C) EBs were labeled with antibodies against LHX2 (dI1, green), FOXD3 (dI2, white), ISL1 (dI3, red) and DAPI (all nuclei, blue). (D, E) Quantification of EBs formed from either d4 (D) or d10 (E) NMPs, suggests that dI1s form robustly from the BMP4+1 condition for both d4 and d10 NMPs. In contrast, dI3s form most robustly from BMP4+4 condition for d4 NMPs and the BMP4+1 condition for d10 NMPs. Probability of similarity between control and experimental groups: * p<0.05, ** p<0.005, *** p<0.0005; two-way ANOVA.

Journal: bioRxiv

Article Title: In vivo human embryonic spinal cord atlas validates stem cell–derived human dorsal interneurons and reveals ASD spinal signatures

doi: 10.64898/2025.12.22.696129

Figure Lengend Snippet: (A, B) EBs were generated at day 0 after either d4 (A) or d10 (B) NMP formation and then treated with RA for 10 days (control). In the experimental samples, BMPs were added at day 1 (BMP4+1), day 2 (BMP4+2) or day 4 (BMP4+4) after EB formation, until day 10. Samples were further differentiated for 6 days and then processed for immunohistochemistry (IHC). (C) EBs were labeled with antibodies against LHX2 (dI1, green), FOXD3 (dI2, white), ISL1 (dI3, red) and DAPI (all nuclei, blue). (D, E) Quantification of EBs formed from either d4 (D) or d10 (E) NMPs, suggests that dI1s form robustly from the BMP4+1 condition for both d4 and d10 NMPs. In contrast, dI3s form most robustly from BMP4+4 condition for d4 NMPs and the BMP4+1 condition for d10 NMPs. Probability of similarity between control and experimental groups: * p<0.05, ** p<0.005, *** p<0.0005; two-way ANOVA.

Article Snippet: The following antibodies were used: LHX2 (mouse; DSHB, PCRP-LHX2-1C11; 1:50), FOXD3 (guinea pig, a gift from Thomas Mueller, Germany, 1:10,000), ISL1 (goat; R&D systems, AF1837; 1:500), PAX2 (rabbit; Invitrogen, 71-6000; 1:500), LMX1B (guinea pig; Thomas Muller Lab), LHX1/5 (mouse; DSHB, 4F2; 1:50) for 12–14 hours (overnight) at 2–8°C in a humidified chamber.

Techniques: Generated, Control, Immunohistochemistry, Labeling