Journal: STAR Protocols

Article Title: Protocol for inducing dorsal spinal sensory interneurons from mouse embryonic stem cell-derived neuromesodermal progenitors

doi: 10.1016/j.xpro.2025.104307

Figure Lengend Snippet: Immunostaining to detect different dI populations (A and C) By day 9, the cultures derived from the RA protocol will contain a mixture of Pax2 + Lhx1/5 + dI4/dI6s and Lmx1b + dI5s (not shown). (B and C) In contrast, the cells derived from the RA+BMP4 contain a mixture of Lhx2 + dI1s and Isl1 + dI3s (B and D). dI2s are present in these cultures but are most effectively detected by scRNA-Seq. Scale bar: 50μm.

Article Snippet: Mouse monoclonal mIgG 2a anti-Lhx2 (1:50) , DSHB , PCRP-LHX2-1C11.

Techniques: Immunostaining, Derivative Assay

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

Journal: bioRxiv

Article Title: Choroid Plexus Hosts the Earliest Detectable Circadian Clock in the Brain

doi: 10.1101/2025.09.09.675224

Figure Lengend Snippet: ( A ) Schematic of the protocol used for recording PER2::LUC bioluminescence from embryonic brain or head slices (left) or from the isolated 4VCP (right). ( B ) Illustration summarizing the finding that the 4VCP is circadian earlier than the SCN in the mouse embryo. ( C ) a : Bright field and PER2::LUC images of the bioluminescence obtained from a PER2::LUC E15.5 brain slice. b : Analysis of the recording showing the period (left) and the circadian power (right). c : Images of immunofluorescence of LHX1 (left) and TTR (right) after brain clearing d : Detrended time series of PER2::LUC bioluminescence from the areas indicated by the white boxes in c. ( D ) a : Bright field and PER2::LUC images of the bioluminescence obtained from a PER2::LUC E13.5 head slice. b : Analysis of the recording showing the period (left) and the circadian power (right). c : PER2::LUC images with white boxes indicating the putative hypothalamus (left) and 4VCP (right). d : Detrended time series of PER2::LUC bioluminescence from the areas indicated by the white boxes in c. ( E ) a : Bright field and PER2::LUC images of the bioluminescence obtained from a PER2::LUC E11.5 head slice. b : Analysis of the recording showing the period (left) and the circadian power (right). c : PER2::LUC images with white boxes indicating the putative hypothalamus (left) and 4VCP (right). d : Detrended time series of PER2::LUC bioluminescence from the areas indicated by the white boxes in c. ( F ) a : PER2::LUC image of the bioluminescence obtained from a PER2::LUC E13.5 isolated 4VCP. b : Analysis of the recording showing the period (left) and the circadian power (right). c : Detrended time series of the recording. Scale bar: 500 µm. Red arrows indicate 4VCP and white arrows indicate SCN.

Article Snippet: Similar steps of wash and blocking were applied before consecutive incubations with LHX1 (AB_531784; 4F2-c, mouse, 1:500, DSHB) and anti-mouse CF350 (SAB4600222, goat, 1:1000, Sigma) as before.

Techniques: Isolation, Slice Preparation, Immunofluorescence