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