Journal: iScience

Article Title: Genetic dissection of the role of Piga and Pgap2 in the embryonic mouse brain

doi: 10.1016/j.isci.2026.116299

Figure Lengend Snippet: Piga in the cerebellum is essential for mammalian survival (A) Percent genotype ratios of En1 Cre/wt ; Piga flox/X females and En1 Cre/wt ; Piga flox/Y males at P0 (Chi-square p = 0.367, n = 109) and P7 (Chi-square p = 0.011, n = 48). H&E stained cerebella (B–G) in wild type (B and E) En1 Cre/wt ; Piga flox/X females (C and F) and En1 Cre/wt ; Piga flox/Y males (D and G) at P0 ( n = 4 animals per genotype, scale bars, 250 μm) and P7 (1–4 animals per genotype, scale bars, 1 mm). (H) Quantification of average cerebellar area per animal at P0 (ANOVA p < 0.001, 4 sections per animals, n = 4 animals each genotype). Immunostaining of SKOR2 marking Purkinje cell progenitors along the ventricular zone in E13 embryo cerebellar hemisphere (I–K scale bars, 100 μm), Calbindin marking Purkinje cells (L–Q), and PAX6 marking granule cell progenitors (R–T) in wildtype (I, L, O, R, n = 5–6 animals), En1 Cre/wt ; Piga flox/X females (J, M, P, S, n = 3–6) and En1 Cre/wt ; Piga flox/Y males (K, N, Q, T, n = 3–4 animals, scale bars, 250 μm). (U) Quantification of PAX6 positive cell per area per animals (ANOVA p = 0.003, average 4 sections per animal). Data represent individual values and the mean ± s.e.m. White arrows indicate the anterior domain of the cerebellar ventricular zone at E13. Black arrows point to ectopic levels of calbindin and white arrows pointing to the posterior domain of the cerebellum at P0.

Article Snippet: Mouse: En1 -Cre; En1 tm2(cre)Wrst /J , The Jackson Laboratory , RRID:IMSR_JAX:007916.

Techniques: Staining, Immunostaining

Journal: iScience

Article Title: Genetic dissection of the role of Piga and Pgap2 in the embryonic mouse brain

doi: 10.1016/j.isci.2026.116299

Figure Lengend Snippet: Pgap2 ablation in the forebrain and the cerebellum does not disrupt brain development (A) X-gal stain of Pgap2 null/wt P21 brain sections (scale bars, 1 mm) in the forebrain cortex (CX), striatum (ST), corpus callosum (CC), and (B) cerebellum (CB, scale bars, 500 μm). Black arrows pointing to the region of enhanced Pgap2 expression in the cortex and cerebellum. (C) Genotype ratios of forebrain ablation Pgap2 animals at P28 ( Emx1 Cre/wt ; Pgap2 flox/wt heterozygous and Emx1 Cre/wt ; Pgap2 flox/null mutant, n = 28, Chi-square p = 0.463). (D–F) Dorsal gross morphological images of wildtype, Emx1 Cre/wt ; Pgap2 flox/wt heterozygous, and Emx1 Cre/wt ; Pgap2 null/flox mutants (scale bars, 1 mm). Measurements of body weights (G, ANOVA p = 0.486), brain weights (H, ANOVA p = 0.823), and brain to body weight ratio (I, ANOVA p = 0.333) in wild type, Emx1 Cre ; Pgap2 flox/wt heterozygous and Emx1 Cre ; Pgap2 null/flox mutant (n = 3–9 animals per genotype). (J) Genotype ratios of cerebellar ablation of Pgap2 animals at P28 ( En1 Cre/wt ; Pgap2 flox/wt heterozygous and En1 Cre/wt ; Pgap2 flox/null mutant, Chi-square p = 0.889). (K–M) Caudal gross morphological images of the cerebellum in wild type, En1 Cre ; Pgap2 flox/wt heterozygous, and En1 Cre/wt ; Pgap2 null/flox mutants. (N–P) Measurements of body weights (N, ANOVA p = 0.614), cerebellar area (O, ANOVA p = 0.523), and cerebellar width (P, ANOVA p = 0.335) in wild type, En1 Cre/wt ; Pgap2 flox/wt heterozygous, and En1 Cre/wt ; Pgap2 flox/null mutants (n = 4–14 animals per genotype). Data represent individual values and the mean ± s.e.m.

Article Snippet: Mouse: En1 -Cre; En1 tm2(cre)Wrst /J , The Jackson Laboratory , RRID:IMSR_JAX:007916.

Techniques: Staining, Expressing, Mutagenesis

Journal: iScience

Article Title: Genetic dissection of the role of Piga and Pgap2 in the embryonic mouse brain

doi: 10.1016/j.isci.2026.116299

Figure Lengend Snippet: Piga in the cerebellum is essential for mammalian survival (A) Percent genotype ratios of En1 Cre/wt ; Piga flox/X females and En1 Cre/wt ; Piga flox/Y males at P0 (Chi-square p = 0.367, n = 109) and P7 (Chi-square p = 0.011, n = 48). H&E stained cerebella (B–G) in wild type (B and E) En1 Cre/wt ; Piga flox/X females (C and F) and En1 Cre/wt ; Piga flox/Y males (D and G) at P0 ( n = 4 animals per genotype, scale bars, 250 μm) and P7 (1–4 animals per genotype, scale bars, 1 mm). (H) Quantification of average cerebellar area per animal at P0 (ANOVA p < 0.001, 4 sections per animals, n = 4 animals each genotype). Immunostaining of SKOR2 marking Purkinje cell progenitors along the ventricular zone in E13 embryo cerebellar hemisphere (I–K scale bars, 100 μm), Calbindin marking Purkinje cells (L–Q), and PAX6 marking granule cell progenitors (R–T) in wildtype (I, L, O, R, n = 5–6 animals), En1 Cre/wt ; Piga flox/X females (J, M, P, S, n = 3–6) and En1 Cre/wt ; Piga flox/Y males (K, N, Q, T, n = 3–4 animals, scale bars, 250 μm). (U) Quantification of PAX6 positive cell per area per animals (ANOVA p = 0.003, average 4 sections per animal). Data represent individual values and the mean ± s.e.m. White arrows indicate the anterior domain of the cerebellar ventricular zone at E13. Black arrows point to ectopic levels of calbindin and white arrows pointing to the posterior domain of the cerebellum at P0.

Article Snippet: En1 tm2(cre)Wrst/J En1−Cre ( En1-Cre , IMSR_JAX:007916 were genotyped using the En1-Cre specific primer from Jackson Labs.

Techniques: Staining, Immunostaining

Journal: iScience

Article Title: Genetic dissection of the role of Piga and Pgap2 in the embryonic mouse brain

doi: 10.1016/j.isci.2026.116299

Figure Lengend Snippet: Pgap2 ablation in the forebrain and the cerebellum does not disrupt brain development (A) X-gal stain of Pgap2 null/wt P21 brain sections (scale bars, 1 mm) in the forebrain cortex (CX), striatum (ST), corpus callosum (CC), and (B) cerebellum (CB, scale bars, 500 μm). Black arrows pointing to the region of enhanced Pgap2 expression in the cortex and cerebellum. (C) Genotype ratios of forebrain ablation Pgap2 animals at P28 ( Emx1 Cre/wt ; Pgap2 flox/wt heterozygous and Emx1 Cre/wt ; Pgap2 flox/null mutant, n = 28, Chi-square p = 0.463). (D–F) Dorsal gross morphological images of wildtype, Emx1 Cre/wt ; Pgap2 flox/wt heterozygous, and Emx1 Cre/wt ; Pgap2 null/flox mutants (scale bars, 1 mm). Measurements of body weights (G, ANOVA p = 0.486), brain weights (H, ANOVA p = 0.823), and brain to body weight ratio (I, ANOVA p = 0.333) in wild type, Emx1 Cre ; Pgap2 flox/wt heterozygous and Emx1 Cre ; Pgap2 null/flox mutant (n = 3–9 animals per genotype). (J) Genotype ratios of cerebellar ablation of Pgap2 animals at P28 ( En1 Cre/wt ; Pgap2 flox/wt heterozygous and En1 Cre/wt ; Pgap2 flox/null mutant, Chi-square p = 0.889). (K–M) Caudal gross morphological images of the cerebellum in wild type, En1 Cre ; Pgap2 flox/wt heterozygous, and En1 Cre/wt ; Pgap2 null/flox mutants. (N–P) Measurements of body weights (N, ANOVA p = 0.614), cerebellar area (O, ANOVA p = 0.523), and cerebellar width (P, ANOVA p = 0.335) in wild type, En1 Cre/wt ; Pgap2 flox/wt heterozygous, and En1 Cre/wt ; Pgap2 flox/null mutants (n = 4–14 animals per genotype). Data represent individual values and the mean ± s.e.m.

Article Snippet: En1 tm2(cre)Wrst/J En1−Cre ( En1-Cre , IMSR_JAX:007916 were genotyped using the En1-Cre specific primer from Jackson Labs.

Techniques: Staining, Expressing, Mutagenesis

Journal: Scientific Reports

Article Title: Region-specific roles of stress-activated protein kinase MKK7 in the developing and maturing murine brain

doi: 10.1038/s41598-025-05388-0

Figure Lengend Snippet: Construction of brain region-specific Mkk7 cKO mice and phenotypes of Mkk7 flox/flox En1-Cre mice. ( A ) Scheme of mating of Cre recombinase-expressing mice with ROSA-tdTomato mice. When recombination occurs due to Cre recombinase expressed from the region-specific promoter, tdTomato is expressed. ( B ) Macroscopic images of brains (left) and spinal cords (right) from mice of the indicated genotypes. ( C ) Cresyl violet staining of sagittal sections of control and Mkk7 flox/flox En1-Cre brains at E18.5. ( D ) Tyrosine hydroxylase (TH) staining of sagittal sections of control and Mkk7 flox/flox En1-Cre brains at E18.5. ( E ) Quantification of the: (i) ratio of the brain lateral ventricle area relative to the whole brain area; (ii) ratio of the cerebellum area relative to the whole brain area; (iii) midbrain thickness; (iv) ratio of the subpallium area relative to the whole brain area for the brains in (C); (v) TH-positive area of the mutant brain relative to control brain as shown in (D). n.s., not significant.

Article Snippet: Mkk7 flox/flox mice were crossed to Hoxb8-Cre (kindly provided by Dr. Zeilhofer, University of Zurich) , En1-Cre (The Jackson Laboratory) , or Emx1-Cre (Riken) transgenic mice expressing Cre recombinase under the control of the Hoxb8 , En1 , or Emx1 promotor, respectively.

Techniques: Expressing, Staining, Control, Mutagenesis

Journal: bioRxiv

Article Title: Development of a protocol utilizing single-cell analysis for the differentiation of human iPSCs into SOX6+ midbrain dopaminergic neurons

doi: 10.1101/2025.06.18.660098

Figure Lengend Snippet: (A) Schematic depiction of the protocol for differentiating mDA neurons, illustrating the application of pharmacological compounds and growth factors. Representative immunocytochemical images and quantification from both original protocol and new protocol showing (B, E) efficacies of midbrain floorplate patterning at DIV11, showing co-localizations of LMX1+/EN1+ and SOX6+/OTX2+; and EN1+/SOX6+/TH+ neurons at DIV 16 (C, F) and 28 (D, G). White scale bars = 50, 100 µm as indicated (n = 3 independent experiments, error bars are SEM, ns = not significant; * = p<0.05; ** = p<0.01; *** = p<0.001; **** = p<0.0001). (H) Calcium imaging recordings of spontaneous and KCl-induced activities on DIV 28 and 42, presenting the total number of calcium events per neuron recorded, and the percentages of cells responding to KCl-induced depolarization. Metrics of activities, including amplitude, rise time, and fall time of calcium signals, are depicted in the subsequent bar plots.

Article Snippet: The primary antibodies were used as follows: CORIN (rat, 1:1,000, R&D Systems, MAB2209), DCX (goat, 1:500, SantaCruz, sc-8066), EN1 (mouse 1:50, DSHB, 4G11), FOXA2 (goat, 1:500, R&D Systems, AF2400), GIRK2 (rabbit, 1:400, Alomone, APC006), LMX1A (rabbit, 1:4,000, Millipore, AB10533), LMO3 (goat, 1:200, SantaCruz, sc-82647), MAP2 (mouse 1:1,000, Sigma, M4403), NURR1 (rabbit, 1:500, SantaCruz, sc-990), OTX2 (goat, 1:1,000, R&D Systems, AF1979), TH (rabbit, 1:1,000, Millipore, AB152), TH (mouse, 1:500, ImmunoStar, 22941), and TH (sheep, 1:500, Novus, NB300).

Techniques: Imaging