cas9 rgn null allele pterem1 impc bay transgenic mice  (International Mouse Phenotyping Consortium)

Journal: Development (Cambridge, England)

Article Title: MEIOC prevents continued mitotic cycling and promotes meiotic entry during mouse oogenesis

doi: 10.1242/dev.205037

Figure Lengend Snippet: Meioc knockout oogenic cells continue to mitotically cycle when wild-type oogenic cells have entered meiosis. (A) Schematic of EdU labeling experiment in E16.5 ovaries. (B) Fluorescent labeling of exemplary EdU in DDX4-positive oogenic cells (arrows) from wild-type, Stra8 knockout, Meioc knockout and Meioc Stra8 double knockout (dKO) ovaries at E16.5. Meioc Stra8 double knockout panel also appears in Fig. 3C . Scale bar: 20 µm. (C) Percentage of DDX4-positive oogenic cells that are positive for EdU in E16.5 ovaries. Error bars represent 95% confidence interval. Numbers on bars represent the total number of DDX4-positive cells quantified. Data were collected from five wild-type embryos, five Stra8 knockout embryos, three Meioc knockout embryos and three Meioc Stra8 double knockout embryos. (D) Log 2 -fold change-based percentile ranks of genes associated with cell cycle phases for wild type versus Stra8 knockout and wild type versus Meioc knockout from whole ovary RNA-seq data. Dot represents the median; whiskers represent the interquartile range. The numbers of genes per cell cycle phase are in parentheses. *adj. P <0.05; **adj. P <0.01; ****adj. P <0.0001; n.s., not significant. See Table S1 for statistical details.

Article Snippet: Mice carrying the Meioc null allele Meioc tm1.1Dcp (RRID:IMSR_JAX:031054; ) and the Stra8 null allele Stra8 tm1Dcp (RRID:IMSR_JAX:023805; ) were backcrossed to C57BL/6N (B6N) from Taconic Biosciences for at least 10 generations.

Techniques: Knock-Out, Labeling, Double Knockout, RNA Sequencing

Journal: Development (Cambridge, England)

Article Title: MEIOC prevents continued mitotic cycling and promotes meiotic entry during mouse oogenesis

doi: 10.1242/dev.205037

Figure Lengend Snippet: CCNA2 protein expression is downregulated during the transition from mitosis to meiosis by MEIOC and STRA8. (A) Fluorescent labeling of CCNA2 in DDX4-positive oogenic cells in mitotic stages (E12.5), transitioning from mitosis to meiosis (E14.5) and in meiotic stages (E16.5). Arrows and arrowheads mark EdU-positive and -negative oogenic cells, respectively. Scale bar: 20 µm. (B) Quantification of CCNA2 intensity in EdU-positive and -negative oogenic cells. Data for EdU-positive cells represents 161 cells from three embryos at E12.5, 240 cells from five embryos at E14.5 and 190 cells from five embryos at E16.5. Data for EdU-negative cells represents 448 cells from three embryos at E12.5, 925 cells from five embryos at E14.5 and 2492 cells from five embryos at E16.5. Dot represents the median; whiskers represent the interquartile range. (C) Labeling of CCNA2 in DDX4-positive oogenic cells from wild-type, Stra8 knockout, Meioc knockout and Meioc Stra8 double knockout (dKO) ovaries at E16.5. Arrows and arrowheads mark EdU-positive and -negative oogenic cells, respectively. Meioc Stra8 double knockout panel also appears in Fig. 1B . Scale bar: 20 µm. (D) Quantification of CCNA2 intensity in EdU-positive and -negative oogenic cells. Data for EdU-positive cells represents 190 cells from five wild-type embryos, 35 cells from five Stra8 knockout embryos, 144 cells from three Meioc knockout embryos and 124 cells from three Meioc Stra8 double knockout embryos. Data for EdU-negative cells represents 2492 cells from five wild-type embryos, 579 cells from five Stra8 knockout embryos, 909 cells from three Meioc knockout embryos and 296 cells from three Meioc Stra8 double knockout embryos. Dot represents the median; whiskers represent the interquartile range. *adj. P <0.05; **adj. P <0.01; ****adj. P <0.0001. See Table S4 for statistical details.

Article Snippet: Mice carrying the Meioc null allele Meioc tm1.1Dcp (RRID:IMSR_JAX:031054; ) and the Stra8 null allele Stra8 tm1Dcp (RRID:IMSR_JAX:023805; ) were backcrossed to C57BL/6N (B6N) from Taconic Biosciences for at least 10 generations.

Techniques: Expressing, Labeling, Knock-Out, Double Knockout

Journal: Development (Cambridge, England)

Article Title: MEIOC prevents continued mitotic cycling and promotes meiotic entry during mouse oogenesis

doi: 10.1242/dev.205037

Figure Lengend Snippet: MEIOC upregulates Meiosin expression and activates the STRA8-MEIOSIN transcription factor to support meiotic entry. (A) Volcano plot of differential expression results from re-analysis of RNA-seq data generated from E14.5 wild-type and Meioc knockout whole ovaries. (B) Venn diagrams comparing gene sets. STRA8-upregulated genes are derived from re-analysis of RNA-seq data generated from E14.5 wild-type and Stra8 knockout whole ovaries. Boxes highlight select Gene Ontology terms and associated genes enriched within each gene set. Meiosin is not yet annotated as a ‘meiotic cell cycle’ gene and is highlighted separately. (C) Percentage of STRA8-activated genes among all expressed genes, STRA8-upregulated genes and MEIOC-upregulated genes in E14.5 ovaries. STRA8-activated genes were defined as those that are upregulated by STRA8 in E14.5 ovaries and bound at the promoter by STRA8 in preleptotene spermatocytes, as defined by . (D) Left: correlation between MEIOC and STRA8 RNA-seq analyses of whole ovaries at E14.5. Analysis is restricted to STRA8-upregulated and -downregulated genes (adjusted P< 0.05). P -value represents the probability that Spearman rho does not equal 0. β Stra8 represents slope in the linear model. Right: distribution of correlations for gene sets obtained by random sampling of genes expressed in MEIOC and STRA8 RNA-seq analyses. P -value represents that probability of obtaining an equal or larger correlation by random sampling. (E) Percentage of meiosis-associated genes upregulated by MEIOC and STRA8 in E14.5 ovaries that are also activated by STRA8 in preleptotene spermatocytes, as reported by . (F) Log 2 -fold change induced by MEIOC and STRA8 in meiosis-associated genes that are statistically upregulated by MEIOC only. **adj. P <0.01; ****adj. P <0.0001. See Table S5 for statistical details.

Article Snippet: Mice carrying the Meioc null allele Meioc tm1.1Dcp (RRID:IMSR_JAX:031054; ) and the Stra8 null allele Stra8 tm1Dcp (RRID:IMSR_JAX:023805; ) were backcrossed to C57BL/6N (B6N) from Taconic Biosciences for at least 10 generations.

Techniques: Expressing, Quantitative Proteomics, RNA Sequencing, Generated, Knock-Out, Derivative Assay, Sampling

Journal: Development (Cambridge, England)

Article Title: MEIOC prevents continued mitotic cycling and promotes meiotic entry during mouse oogenesis

doi: 10.1242/dev.205037

Figure Lengend Snippet: MEIOC collaborates with STRA8-MEIOSIN to rewire the cell cycle program as oogenic cells transition from mitosis to meiosis. Purple highlights the previously unreported regulation discovered in this study. Pointed and blunt black arrows indicate active regulation; pointed and blunt gray arrows indicate regulation that is inactive due to upstream activity. MEIOC inhibits CCNA2 to prevent continued mitotic cycling and activates the STRA8-MEIOSIN transcriptional regulator to drive meiotic entry. In the absence of MEIOC, oogenic cells undergo additional mitotic cycles. Eventually, at least some oogenic cells stochastically express sufficient Meiosin levels independently of MEIOC to promote meiotic entry. In the absence of STRA8, oogenic cells arrest at the G1/S phase transition without entering meiosis, although some oogenic cells do slip into an early S phase.

Article Snippet: Mice carrying the Meioc null allele Meioc tm1.1Dcp (RRID:IMSR_JAX:031054; ) and the Stra8 null allele Stra8 tm1Dcp (RRID:IMSR_JAX:023805; ) were backcrossed to C57BL/6N (B6N) from Taconic Biosciences for at least 10 generations.

Techniques: Activity Assay, Sublimation

Journal: Cell Death Discovery

Article Title: MKK4 and MKK7 control degeneration of retinal ganglion cell somas and axons after glaucoma-relevant injury

doi: 10.1038/s41420-025-02842-w

Figure Lengend Snippet: A Longitudinal intraocular pressure (IOP) measurements from D2 and D2. Ddit3/Jun −/− eyes ( n listed respectively here and throughout figure legends) at 5 M ( n = 60, 63), 9 M ( n = 60, 63), 10.5 M ( n = 66, 63), and 12 M ( n = 62, 61). Both genotype groups had significantly elevated IOPs at 9 M, 10.5 M, and 12 M compared to 5 M (* P < 0.001). At these timepoints, D2. Ddit3/Jun −/− eyes did not have a statistically significant reduction in IOP compared to WT, although D2. Ddit3/Jun −/− eyes had slightly higher IOPs at 9 M of age compared to D2 (* P = 0.004). Two-way ANOVA, Holm-Sidak’s post hoc . B Examples of optic nerve cross sections with no or early (noe) and severe (sev) glaucomatous damage from D2 and D2. Ddit3/Jun −/− mice and percentages of optic nerves with noe ( n = 23, 16), moderate (mod; n = 3, 11) and sev ( n = 31, 27) glaucomatous damage. Ddit3/Jun deletion did not afford protection to RGC axons and in fact slightly worsened axonal degeneration (* P = 0.049). Chi square test. C Representative retinal flat mounts immunoassayed for RBPMS and quantification of RBPMS+ cells from 12 M D2 and D2. Ddit3/Jun −/− retinas with corresponding noe ( n = 6, 6) or sev ( n = 6, 6) optic nerves. Sev D2. Ddit3/Jun −/− retinas had 77.0 ± 3.1% improved RGC survival compared to D2 controls (* P < 0.001). RBPMS+ cells/mm 2 ± SEM for D2 and D2. Ddit3/Jun −/− respectively: Noe: 2899.6 ± 111.0, 2765.4 ± 49.8; Sev: 226.7 ± 20.8, 2151.5 ± 83.0. Two-way ANOVA, Holm-Sidak post-hoc . Scale bars, 50 μm.

Article Snippet: Ddit3 null alleles [ ] (Jackson Laboratory, Stock# 005530), floxed alleles of Jun [ ] ( Jun fl ), and the Six3-cre transgene [ ] (Jackson Laboratory, Stock# 019755) were backcrossed >10 times to both the C57BL/6 J genetic background (>99% C57BL/6J) and the DBA/2J background (>99% DBA/2J).

Techniques:

Journal: Cell Death Discovery

Article Title: MKK4 and MKK7 control degeneration of retinal ganglion cell somas and axons after glaucoma-relevant injury

doi: 10.1038/s41420-025-02842-w

Figure Lengend Snippet: A Representative pattern electroretinography (PERG) traces and amplitude quantification ( B ) from D2. Gpnmb + , D2, D2. Ddit3 −/− , D2. Jun −/− , and D2. Ddit3/Jun −/− mice at 5 M ( n = 20, 34, 38, 34, 39), 9 M ( n = 22, 34, 38, 39, 36), and 12 M ( n = 21, 34, 36, 35, 35). Normotensive D2. Gpnmb + mice did not have significant decline in PERG amplitude at 9 M ( P = 0.220), but had a slight but significant decline in PERG amplitude by 12 M compared to 5 M (* P = 0.001), Ocular hypertensive mice of all genotype groups had significant PERG amplitude decline at 9 M and 12 M compared to 5 M (* P < 0.001). At 9 M and 12 M, each ocular hypertensive group’s PERG amplitude was significantly lower than normotensive D2, Gpnmb + controls (* P < 0.001). No increase in PERG amplitude was observed between D2 and D2. Ddit3 −/− , D2. Jun −/− , or D2. Ddit3/Jun −/− groups at any timepoint measured. Scale bar: Y: 5 μV, X: 100 ms. Mixed effects analysis, Holm-Sidak’s post hoc . C Quantification of full-field ERG a-wave and b-wave ( D ) amplitudes in D2. Gpnmb + , D2, D2. Ddit3 −/− , D2. Jun −/− , and D2. Ddit3/Jun −/− eyes. By 12 M ( n = 21, 36, 35, 33, 37), each ocular hypertensive group had a slight but significant decline of electroretinography (ERG) a and b-wave amplitudes compared to 5 M ( n = 20, 38, 30, 34, 30) (* P < 0.001), but not nearly to the same extent as PERG amplitude decline ( A ). Two-way ANOVA, Holm-Sidak’s post hoc . Percentage of PERG and ERG amplitude declines at 9 and 12 M are listed for each group in Table . E PERG amplitude quantifications from 12 M D2 and D2. Ddit3/Jun -/- retinas with noe ( n = 6, 7) and sev ( n = 8, 17) optic nerve damage. Neither genotype nor optic nerve damage level influenced PERG amplitude ( P > 0.05, Two-way ANOVA). Of note, PERG amplitudes were significantly reduced compared with 12 M D2. Gpnmb + ( n = 21, 7.2 ± 0.6) regardless of genotype or level of axonal damage (* P < 0.001, One-way ANOVA, Holm-Sidak’s post hoc ). PERG amplitude (μV) ± SEM from D2 and D2. Ddit3/Jun −/− retinas, respectively: noe: 2.0 ± 0.4, 1.8 ± 0.2; sev: 2.3 ± 0.3; 1.9 ± 0.2. Scale bar: Y: 5 μV, X: 100 ms. F High-resolution images of retinal flat mounts immunoassayed for RBPMS (scale bar, 50μm) and quantification of average RGC soma size from D2 and D2. Ddit3/Jun −/− retinas with noe ( n = 7, 5) and sev ( n = 6, 6) glaucomatous damage. Both genotype groups had significant reductions in RGC soma size in sev glaucoma compared to respective noe controls (* P < 0.001). While D2. Ddit3/Jun −/− noe retinas had slightly smaller RGCs (* P = 0.044), Ddit3/Jun deletion did not attenuate RGC soma shrinkage in sev retinas. Soma size (μm 2 ) ± SEM from D2 and D2. Ddit3/Jun −/− retinas, respectively: noe: 143.1 ± 3.9, 131.7 ± 2.9; sev: 87.6 ± 2.5, 78.3 ± 3.0. Two-way ANOVA, Holm-Sidak’s post hoc .

Article Snippet: Ddit3 null alleles [ ] (Jackson Laboratory, Stock# 005530), floxed alleles of Jun [ ] ( Jun fl ), and the Six3-cre transgene [ ] (Jackson Laboratory, Stock# 019755) were backcrossed >10 times to both the C57BL/6 J genetic background (>99% C57BL/6J) and the DBA/2J background (>99% DBA/2J).

Techniques:

Journal: Cell Death Discovery

Article Title: MKK4 and MKK7 control degeneration of retinal ganglion cell somas and axons after glaucoma-relevant injury

doi: 10.1038/s41420-025-02842-w

Figure Lengend Snippet: A Representative sections and quantification of IPL length from B6, B6. Ddit3/Jun −/− , and B6. Mkk4/7 −/− retinas 35 days post-Sham ( n = 6, 5, 7) or CONC ( n = 6, 5, 8) procedures. B6 IPL length significantly decreased 35 days post-CONC (by 22.7 ± 11%, * P = 0.030), while B6. Ddit3/Jun −/− and B6. Mkk4/7 −/− IPL lengths did not significantly change after CONC ( P = 0.883 and P = 0.925, respectively). B6. Mkk4/7 −/− IPLs were significantly longer than B6 IPLs post-CONC (* P = 0.020). Note, in this experiment, Jun fl alleles were recombined from the retina using bilateral intravitreal delivery of AAV2.2-CMV-Cre-GFP. IPL length (µm) ± SEM for B6, B6. Ddit3/Jun −/− and B6. Mkk4/7 −/− mice, respectively: Sham: 50.1 ± 3.6, 51.8 ± 4.3, 52.8 ± 2.8; CONC: 38.7 ± 5.5, 51.0 ± 2.7, 52.4 ± 1.7. Two-way ANOVA, Holm-Sidak’s post hoc . Scale bar, 50 μm. B Representative PERG traces and quantification of PERG amplitudes from B6, B6. Ddit3/Jun −/− , and B6. Mkk4/7 −/− retinas 14 days post- Sham ( n = 33, 17, 20) or CONC ( n = 35, 17, 20) procedures. B6 and B6. Ddit3/Jun −/− PERG amplitudes significantly declined 14 days post-CONC (by 52.6 ± 5.7%, * P < 0.001 and 57.9 ± 7.9%, * P = 0.002, respectively), while B6. Mkk4/7 −/− PERG amplitudes did not significantly decline ( P = 0.108). Two-way ANOVA, Holm-Sidak’s post-hoc . Scale bar: Y: 5 μV, X: 100 ms. C Representative high-resolution images of retinal flat mounts immunoassayed for RBPMS and quantification of RGC soma size from B6. Ddit3/Jun −/− and B6. Mkk4/7 −/− retinas 14 days post-Sham ( n = 8, 9) or CONC ( n = 7, 9) procedures. After CONC, B6. Ddit3/Jun −/− RGC somas were reduced to 65.5 ± 4.6% (* P < 0.001) the size of respective RGCs after Sham procedures. B6. Mkk4/7 −/− RGC soma sizes did not change after CONC compared to Sham ( P = 0.439). B6. Mkk4/7 −/− RGCs were significantly larger compared to B6. Ddit3/Jun −/− (* P < 0.001) RGCs after CONC. Soma size (μm 2 ) ± SEM from B6. Ddit3/Jun −/− and B6. Mkk4/7 −/− mice, respectively: Sham: 137.0 ± 7.3, 155.8 ± 3.2; CONC: 89.7 ± 6.3, 147.5 ± 7.8. Two-way ANOVA, Holm-Sidak’s post hoc . Scale bar, 50 μm.

Article Snippet: Ddit3 null alleles [ ] (Jackson Laboratory, Stock# 005530), floxed alleles of Jun [ ] ( Jun fl ), and the Six3-cre transgene [ ] (Jackson Laboratory, Stock# 019755) were backcrossed >10 times to both the C57BL/6 J genetic background (>99% C57BL/6J) and the DBA/2J background (>99% DBA/2J).

Techniques:

Journal: Nature Communications

Article Title: Targeting Msx2 as a brake in the fusion fate of osteoclasts and an anabolic therapy in pre-clinical models of osteoporosis

doi: 10.1038/s41467-025-61938-0

Figure Lengend Snippet: a BMMs were stimulated by RANKL for 3 days, and immunofluorescence staining of MSX2 and PU.1 in BMMs were captured. Scale bar, 20 μm. b BMMs were stimulated by RANKL for 3 days, and Co-IP was performed to show the interaction between MSX2 and PU.1. c HEK-293T cells were transfected by PU.1-FLAG and MSX2-HA, and the interaction between MSX2 and PU.1 were examined. d Prediction of the E3 ubiquitination ligase of PU.1 using Ubibrowser. e HEK-293T cells were transfected as indicated and then treated by MG132 for 6 h. The ubiquitination level of PU.1 was detected by Co-IP. f The FBXW7 CPD consensus motif in mouse PU.1. g Molecular docking of MSX2 and PU.1 complex. h HEK-293T cells were transfected as indicated and then treated by MG132 for 6 h. The PU.1 ubiquitination levels were detected by Co-IP. i Schematic of the protective effect of MSX2 on PU.1 ubiquitination degradation. All experiments were repeated 3 times independently, and the representative images were shown ( a–c , e , and h ).

Article Snippet: Mice carrying Fbxw7 conditional null alleles ( Fbxw7 f/f , NM-CKO-200327) were from Shanghai Model Organisms (Shanghai, China).

Techniques: Immunofluorescence, Staining, Co-Immunoprecipitation Assay, Transfection, Ubiquitin Proteomics

Journal: Nature Communications

Article Title: Targeting Msx2 as a brake in the fusion fate of osteoclasts and an anabolic therapy in pre-clinical models of osteoporosis

doi: 10.1038/s41467-025-61938-0

Figure Lengend Snippet: a Representative 3D Micro-CT images of distal femurs from 3-month-old female Msx2 f/f and Msx2 cKO and Msx2 Fbxw7 dKO mice. b Bone morphology parameters of distal femurs from Msx2 f/f ( n = 8), Msx2 cKO ( n = 8), and Msx2 Fbxw7 dKO ( n = 8) mice. c Serum CTX-1 level of Msx2 f/f , Msx2 cKO, and Msx2 Fbxw7 dKO mice ( n = 5). d Representative TRAP staining images of distal femurs from Msx2 f/f , Msx2 cKO, and Msx2 Fbxw7 dKO mice. Scale bar, 100 μm. e Quantification of Mu.Oc.N/BS in ( d ) ( n = 5). f OCN immunofluorescence staining of distal femurs. Scale bar, 50 μm. g N.OCN + cells/BS was measured ( n = 5). h Representative images of EMCN and CD31 immunofluorescence staining of distal femurs. Scale bar, 50 μm. i CD31 hi EMCN hi .Ar was measured ( n = 5). j Representative images of PDGF-BB and TRAP immunofluorescence staining. Scale bar, 50 μm. k Quantification of PDGF-BB + TRAP + cell numbers/ BS ( n = 5). Data are presented as mean ± SD, and p -values were calculated using one-way ANOVA with Tukey’s multiple comparisons.

Article Snippet: Mice carrying Fbxw7 conditional null alleles ( Fbxw7 f/f , NM-CKO-200327) were from Shanghai Model Organisms (Shanghai, China).

Techniques: Micro-CT, Staining, Immunofluorescence

Journal: Nature Communications

Article Title: Targeting Msx2 as a brake in the fusion fate of osteoclasts and an anabolic therapy in pre-clinical models of osteoporosis

doi: 10.1038/s41467-025-61938-0

Figure Lengend Snippet: In wild-type myeloid cells, MSX2 binds PU.1 and prevents PU.1 from FBXW7 mediated ubiquitination, which safeguards fusion cell fate of osteoclast and results in bone resorption. Targeting Msx2 promotes PU.1 degradation and acts as a “brake” in the progression of the fusion fate of osteoclast, resulting in a larger population of pre-osteoclasts. Pre-osteoclasts secrete PDGF-BB and enhance angiogenesis-mediated bone formation (upper part of Fig. 8). Since voluminous prior studies have reported that MSX2 is important in regulating Wnt signaling and bone formation in concert with osteogenic transcription factors, myeloid-specific targeting MSX2 is necessary to achieve net anabolic effects (lower part of Fig. 8). The schematic diagram was designed by Figdraw.

Article Snippet: Mice carrying Fbxw7 conditional null alleles ( Fbxw7 f/f , NM-CKO-200327) were from Shanghai Model Organisms (Shanghai, China).

Techniques: Ubiquitin Proteomics