Journal: bioRxiv

Article Title: Elongation of Very Long Chain Fatty Acids Like- 3 ( Elovl3 ) is activated by ZHX2 and is a regulator of cell cycle progression

doi: 10.1101/2022.09.02.506374

Figure Lengend Snippet: RNA was prepared from the livers of adult male (A) BALB/cByJ (cByJ; Zhx2+) and BALB/cJ (cJ; Zhx2-) mice and (B-D) BL/6 mice that express ( Zhx2 f/f ) or do not express Zhx2 in hepatocytes ( Zhx2 Δ hep ). Elovl3 (A,C), Zhx2 (B) and AFP (D) levels were measured by RT-qPCR and normalized to Sfrs4 (A) or ribosomal protein L30 (B,C,D) mRNA levels. Mean (SD), analyzed by Student’s t-test, (n=3-5/group). *, p<0.05; **, p<0.01; *** p<0.001.

Article Snippet: Briefly, adult male mouse livers from Zhx2 WT and Zhx2 KO mice were harvested and processed for ChIP with rabbit anti-Zhx2 antibodies (Bethyl labs) or control IgG using the Magna ChIP HiSense kit (Millipore).

Techniques: Quantitative RT-PCR

Journal: bioRxiv

Article Title: Elongation of Very Long Chain Fatty Acids Like- 3 ( Elovl3 ) is activated by ZHX2 and is a regulator of cell cycle progression

doi: 10.1101/2022.09.02.506374

Figure Lengend Snippet: (A, B) . Human hepatoma HepG2 cells were transiently transfected with a pcDNA vector containing flag-tagged Zhx2 (Zhx2-flag) or pcDNA empty vector control (pcDNA). After 48 hours, cells were harvested and RNA and protein extracts were prepared. (A) Western blots were probed using anti-Flag antibodies and reprobed with anti-actin antibodies to confirm equal lysate loading. (B) RT-qPCR analysis was performed to measure Elovl3 and AFP mRNA levels normalized to GAPDH . Data is representative of three separate experiments. (C) . Adult male liver samples from Zhx WT and Zhx2 KO mice were analyzed by ChIP using anti-Zhx2 antibodies or control IgG and primers that generate amplicons from the Elovl3 promoter, 3’ DHS site or 5’ DHS site. For all three regions, levels of IgG in Zhx2 WT and Zhx2 KO mice were set to 1. Data are from three separate experiments. **, p<0.01 (D). DNAse Hypersensitive sites from ENCODE adult mouse liver flanking the mouse Elovl3 gene are shown as yellow peaks (designated by arrows) in analysis using the University of California – Santa Cruz (UCSC) Genome Browser.

Article Snippet: Briefly, adult male mouse livers from Zhx2 WT and Zhx2 KO mice were harvested and processed for ChIP with rabbit anti-Zhx2 antibodies (Bethyl labs) or control IgG using the Magna ChIP HiSense kit (Millipore).

Techniques: Transfection, Plasmid Preparation, Control, Western Blot, Quantitative RT-PCR

Journal: bioRxiv

Article Title: Elongation of Very Long Chain Fatty Acids Like- 3 ( Elovl3 ) is activated by ZHX2 and is a regulator of cell cycle progression

doi: 10.1101/2022.09.02.506374

Figure Lengend Snippet: Liver mRNA was prepared from male and female B6C3F1 mice at embryonic day 17.5 (e17.5) and postnatal day 1 (p1), p7, p14, p21, p28 and p56. RT-qPCR analysis was performed to assess Zhx2 (panels A and C) and Elovl3 (panels B and D) mRNA levels in male (panels A and B) and female (panels C and D) mice, which were normalized to L30 . Mean (SD), analyzed by ANOVA with Dunnett’s test for multiple comparisons; each timepoint compared to e17.5 expression. n= 4-10 mice/group. *, <0.05; ***, <0.001; ****, <0.0001.

Article Snippet: Briefly, adult male mouse livers from Zhx2 WT and Zhx2 KO mice were harvested and processed for ChIP with rabbit anti-Zhx2 antibodies (Bethyl labs) or control IgG using the Magna ChIP HiSense kit (Millipore).

Techniques: Quantitative RT-PCR, Expressing