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

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

Journal: Frontiers in Molecular Biosciences

Article Title: Hepatocyte-Specific Knock-Out of Nfib Aggravates Hepatocellular Tumorigenesis via Enhancing Urea Cycle

doi: 10.3389/fmolb.2022.875324

Figure Lengend Snippet: Generation of hepatocyte-specific NFIB-knockout mice (Nfib f/f Alb-Cre). (A) NFIB gene knockout targeting vector strategy design diagram. (B) Genotyping of Nfib loxp/loxp Alb-cre (Nfib f/f Alb-Cre) mice using the tail DNA. The above panel shows the homozygous loxp mice. The wild-type (WT) allele yields an amplicon of 162 bp, while the floxed allele yields an amplicon of 229 bp. The panel below shows the Alb-Cre positive mice, with an amplicon of 390 bp. (C) IHC analysis of NFIB protein in the livers of WT and Nfib f/f Alb-Cre mice. (D) H&E staining showing the histology of the livers of WT and Nfib f/f Alb-Cre mice. All scale bars = 100 μm. Original magnification×20. IHC, immunohistochemistry.

Article Snippet: Mice with homozygous conditional-null alleles of NFIB (Nfib f/f ) were generated with the help of the Cyagen Biosciences (Guangzhou, China).

Techniques: Knock-Out, Gene Knockout, Plasmid Preparation, Amplification, Staining, Immunohistochemistry

Journal: The Journal of Clinical Endocrinology and Metabolism

Article Title: Alterations in Sod2-Induced Oxidative Stress Affect Endocrine Cancer Progression

doi: 10.1210/jc.2018-01039

Figure Lengend Snippet: OS pathways in human endocrine neoplasia. Oxidative pathway and Sod2 expression was analyzed in different endocrine tumors. (A) Deregulation of OS genes in FTC compared with follicular adenoma, analyzed from microarray data. Significantly altered genes above cutoff (P < 0.01) are indicated by red circles; those below cutoff are indicated by gray circles; Sod2 is indicated by an arrow. (B) Expression of Sod2 in PTC compared with normal tissue (left), and matched primary tissue vs metastatic tissue (right). (C) Analysis of Sod2 expression in ATC compared with poorly differentiated thyroid cancer. (D) Kaplan-Meier survival curves of data from patients with ATC, based on low (red line) and high (green line) expression. (E) Kaplan-Meier survival curves of data from patients with ACC based on low (red line) and high (green line) expression. P value was calculated by log-rank test in survival analysis. Sources of microarray data for these analyses are described in Methods. **P < 0.01. ns, not significant; RSEM, RNA-sequencing by expectation-maximization.

Article Snippet: To create mice lacking one copy of Sod2 ( i.e. , haploinsufficiency), these mice were crossed independently with animals heterozygous for a null allele of Sod2 (stock no. 002973; Jackson Laboratories) ( 32 ).

Techniques: Expressing, Microarray, RNA Sequencing

Journal: The Journal of Clinical Endocrinology and Metabolism

Article Title: Alterations in Sod2-Induced Oxidative Stress Affect Endocrine Cancer Progression

doi: 10.1210/jc.2018-01039

Figure Lengend Snippet: Deregulation of OS genes in murine tumor progression models. Microarray data comparing Pten-, R1a-, and DRP-TpoKO tumors. Oxidative pathways genes were analyzed in (A) Pten-TpoKO tumors, (B) R1a-TpoKO tumors, and (C) DRP-TpoKO tumors. Significantly altered genes above cutoff (P < 0.01) are indicated by red circles; those below the cutoff are indicated by gray circles; Sod2 is indicated by the arrow in panels A–C. Wild-type Cre-negative littermates were used for comparison for each model. (D) Expression of Sod2 in each tumor model compared with wild-type control. Dotted line represents the fold change of Cre-negative control tissue for each group. **P < 0.01.

Article Snippet: To create mice lacking one copy of Sod2 ( i.e. , haploinsufficiency), these mice were crossed independently with animals heterozygous for a null allele of Sod2 (stock no. 002973; Jackson Laboratories) ( 32 ).

Techniques: Microarray, Comparison, Expressing, Control, Negative Control

Journal: The Journal of Clinical Endocrinology and Metabolism

Article Title: Alterations in Sod2-Induced Oxidative Stress Affect Endocrine Cancer Progression

doi: 10.1210/jc.2018-01039

Figure Lengend Snippet: Sod2 overexpression increases tumor aggressiveness in a Pten KO mouse model of FA. (A, top) Three-dimensional rendering of ultrasonographic images of Pten-TpoKO with Sod2-wt, Sod2+/− and Sod2-Tg at 12 months. (A, bottom) Average thyroid volumes determined by three-dimensional ultrasonography at 3, 6, 9, and 12 mos in Pten-TpoKO mice with Sod2-wt (black line; n = 12), Sod2+/− (red line; n = 22), and Sod2-Tg (green line; n = 18). (B) The incidence of thyroid carcinoma in Pten-TpoKO mice with Sod2-wt, Sod2+/−, and Sod2-Tg. (C, top) Representative ×40 images of Ki67 staining in thyroid tumors of mice at 12 mos of age. (C, bottom) Quantification of proliferation represented as percent DAB to nuclear ratio. Graphs present mean data ± SD. *P ≤ 0.05. DAB, diaminobenzidine.

Article Snippet: To create mice lacking one copy of Sod2 ( i.e. , haploinsufficiency), these mice were crossed independently with animals heterozygous for a null allele of Sod2 (stock no. 002973; Jackson Laboratories) ( 32 ).

Techniques: Over Expression, Staining

Journal: The Journal of Clinical Endocrinology and Metabolism

Article Title: Alterations in Sod2-Induced Oxidative Stress Affect Endocrine Cancer Progression

doi: 10.1210/jc.2018-01039

Figure Lengend Snippet: Sod2 deficiency induces tumor growth in a dual Pten/Prkar1a KO mouse model of metastatic FTC. (A, top) Three-dimensional rendering of ultrasonographic images of thyroid glands in DRP-TpoKO with Sod2-wt, Sod2+/−, and Sod2-Tg mice at 12 mos. (A, bottom) Average thyroid volumes determined by three-dimensional ultrasonography at 3, 6, 9, and 12 mos in DRP-TpoKO mice with Sod2-wt (black line; n = 16), Sod2+/− (red line; n = 12), and Sod2-Tg (green line; n = 20). (B) Kaplan-Meier survival curves of DRP-TpoKO mice with Sod2-wt (black line), Sod2 deficiency (red line), and Sod2 overexpression (green line). (C, top) Representative ×40 images of Ki67 staining in thyroid tumors of mice at 12 mos of age. (C, bottom) Quantification of proliferation represented as percent DAB to nuclear ratio. (D) Percent incidence of lung metastases in DRP-TpoKO mice with Sod2-wt, Sod2+/−, and Sod2-Tg. *P ≤ 0.05; **P ≤ 0.01. DAB, diaminobenzidine.

Article Snippet: To create mice lacking one copy of Sod2 ( i.e. , haploinsufficiency), these mice were crossed independently with animals heterozygous for a null allele of Sod2 (stock no. 002973; Jackson Laboratories) ( 32 ).

Techniques: Over Expression, Staining