Journal: Redox biology

Article Title: SIRT3 improved peroxisomes-mitochondria interplay and prevented cardiac hypertrophy via preserving PEX5 expression.

doi: 10.1016/j.redox.2023.102652

Figure Lengend Snippet: Fig. 1. SIRT3 regulated PEX5 expression level in cardiomyocyte hypertrophy. (A) Venn diagrams showed the number of differentiated expression genes from heart tissue extracted from Ang II induced cardio-hypertrophic mice (WT mice, male, 10–12 weeks old) and Sirt3 knock out mice (male, 28 weeks old). (B) The heat maps showed the alteration of expression level of selected genes according to microarray analysis results. Values of normalized intensity for differentiated genes was there for comparison. (C) qPCR validation result of selected genes at cellular level by silencing SIRT3 in NRCMs. (D) The protein level of PEX5 in heart tissue from SIRT3 KO mice and wild type mice were measured by Western blot. (E) NRCMs were transfected with SIRT3 siRNA with or without PE treatment (Ctrl siRNA was used as negative control). The protein level of SIRT3 and PEX5 was examined by Western blot. (F) NRCMs were infected by Ad SIRT3 with or without PE treatment (Ad GFP was used as negative control). The protein level of SIRT3 and PEX5 was tested by Western blot. All data were showed as the means ± SEM. *P < 0.05. **P < 0.01. ***P < 0.001. n = 3.

Article Snippet: Male Sirt3 knockout mice (Strain# 129-Sirt3tm1.1Fwa/J) and their age-, sex- and background-matched wild-type (WT) control (129/SvlmJ) mice were purchased from Jackson Laboratory (Bar Harbor, ME, USA).

Techniques: Expressing, Knock-Out, Microarray, Comparison, Biomarker Discovery, Western Blot, Transfection, Negative Control, Infection

Journal: Redox biology

Article Title: SIRT3 improved peroxisomes-mitochondria interplay and prevented cardiac hypertrophy via preserving PEX5 expression.

doi: 10.1016/j.redox.2023.102652

Figure Lengend Snippet: Fig. 2. PEX5 was involved in inhibitory effect of SIRT3 on cardiomyocyte hypertrophy. NRCMs were simultaneously infected with Ad SIRT3 and transfected with PEX5 siRNA in settings of PE (100 μM，24h) treatment. Ad GFP was used as negative control to Ad SIRT3 and Ctrl siRNA was used as negative control to PEX5 siRNA (A, C). Or NRCMs were simultaneously treated with SIRT3 inhibitor 3-TYP (50 μM, 12h) and transfected with Flag-PEX5 plasmid in settings of PE (100 μM，24h) treatment. DMSO was used as negative control to 3-TYP and vector was used as negative control to Flag-PEX5 plasmid (B, D). The protein level of hypertrophic marker ANF and β-MHC was detected by Western blot (A, B). Cardiomyocyte surface area was examined by Rhodamine-Phalloidin staining and calculated by High- Content Screening System (C, D). The data were presented as the means ± SEM. *P < 0.05. n = 3.

Article Snippet: Male Sirt3 knockout mice (Strain# 129-Sirt3tm1.1Fwa/J) and their age-, sex- and background-matched wild-type (WT) control (129/SvlmJ) mice were purchased from Jackson Laboratory (Bar Harbor, ME, USA).

Techniques: Infection, Transfection, Negative Control, Plasmid Preparation, Marker, Western Blot, Staining, High Content Screening

Journal: ACS Pharmacology & Translational Science

Article Title: Chemokine Receptor 2 Targeted PET/CT Imaging Distant Metastases in Pancreatic Ductal Adenocarcinoma

doi: 10.1021/acsptsci.3c00303

Figure Lengend Snippet: 68 Ga-DOTA-ECL1i PET/CT is detecting liver metastasis in a KPC mouse model and tissue characterization. (A) 68 Ga-DOTA-ECL1i PET/CT images in 16–18 and 20–23 weeks old KPC mice and 20 weeks old control littermates. (B) 68 Ga-DOTA-ECL1i PET quantification and metastasis/liver ratios. (C) Autoradiographic images and pictures of sliced liver from a 20–23 week-old KPC mouse. (D) H&E of the liver slice revealed two small regions with metastasis (black arrows). H&E and immunofluorescence staining showed CCR2 + cells in liver metastasis. M: liver metastasis. T: primary tumor.

Article Snippet: KPC mice were obtained from the interbreeding of p48-CRE, LSL-KRAS G12D/wt , and p53 flox / flox 1-Cre strains on a mixed 129/SvJae/C57Bl/6 (The Jackson Lab) background.

Techniques: Positron Emission Tomography-Computed Tomography, Control, Immunofluorescence, Staining

Journal: Cell

Article Title: Renal PIEZO2 is an essential regulator of renin

doi: 10.1016/j.cell.2025.11.013

Figure Lengend Snippet: A. Representative Sun1-sfGFP fl/fl ; Pdgfrb CreERT2 mouse kidney. B. UMAP projection of Sun1-sfGFP fl/fl ; Pdgfrb CreERT2 snRNA-seq. C . Multidimensional dot plot of cell type-specific markers used to identify clusters corresponding to distinct Pdgfrb + kidney cell populations in D . Violin plot of Piezo2 . E. Feature plot of Piezo2 expression in UMAP space. F . Violin plot of Piezo1 . G. Feature plot of Piezo1 expression in UMAP space. See also – .

Article Snippet: The following strains of mice were used and maintained in the laboratory on an inbred background: Piezo1 tdTomato ( B6;129-Piezo1 tm1.1Apat/ J ; Jackson Laboratories 029214) and Piezo2 EGFP-IRES-Cre ( B6(SJL)-Piezo2 tm1.1(cre)Apat /J ; Jackson Laboratories 027719).

Techniques: Expressing

Journal: eLife

Article Title: HIPPO signaling resolves embryonic cell fate conflicts during establishment of pluripotency in vivo

doi: 10.7554/elife.42298

Figure Lengend Snippet: Figure 1. ROCK1/2 and nuclear YAP1 repress expression of SOX2. (A) Experimental design: embryos were collected at E2.5 and treated with ROCK inhibitor Y-27632 (ROCKi) or DMSO (control) for 24 hr. (B–B’) Confocal images of apical (PARD6B) and basolateral (CDH1) membrane components in control and ROCKi-treated embryos. As expected, PARD6B and CDH1 are mislocalized to the entire cell membrane of all cells in ROCKi-treated embryos, demonstrating effective ROCK inhibition (n = number of embryos examined). (C–C’) In control embryos, SOX2 is detected only in inside cells, while in ROCKi-treated embryos, SOX2 is detected in inside and outside cells (arrowheads, outside cells; n = embryos). (D) Quantification of ectopic SOX2 detected in outside cells of control and ROCKi-treated embryos (p, student’s t-test, n = embryos). (E) SOX2 and CDX2 staining in outside cells of control and ROCKi-treated embryos. ROCK-inhibitor treatment leads to outside cells with mixed lineage marker expression (CDX2+/SOX2+). (F) Experimental design: embryos were collected at E1.5 and one of two blastomeres injected with mRNAs encoding YAP1CA and GFP. Embryos were cultured for 72 hr, fixed, and then analyzed by immunofluorescence and confocal microscopy. (G) SOX2 is detected non-injected inside cells. SOX2 is not detected in YAP1CA-overexpressing inside cells (arrowheads), n = embryos. (H) Across multiple embryos, all non-injected inside cells express SOX2, whereas the vast majority of YAP1CA-injected inside cells fail to express SOX2. DOI: https://doi.org/10.7554/eLife.42298.003 The following figure supplement is available for figure 1:

Article Snippet: Key resources table Reagent type (species) or resource Designation Source or reference Identifiers Additional information Strain, strain background (Mus musculus) CD-1 Charles River Laboratories RRID:IMSR _CRL:22 Strain, strain background (M. musculus) Sox2tm1.1Lan Smith et al. (2009) PMID:19666824 RRID:IMSR_ JAX:013093 mixed background, Sox2 null refers to recombined allele Strain, strain background (M. musculus) Wwtr1 conditional allele; Wwtr1tm1.1Eno; Wwtr1loxp Xin et al., 2013 Xin et al., 2013 PMID:23918388 MGI:5544289 mixed background, ‘Wwtr1-’ or ‘Wwtr1 D ’ refers to recombined allele Strain, strain background (M. musculus) Yap1 conditional allele; Yap1tm1.1Eno; Yap1loxp Xin et al., 2011 Xin et al., 2011 PMID:22028467 MGI:5446483 mixed background, , ‘Yap1-’ or ‘Yap1 D ’ refers to recombined allele Strain, strain background (M. musculus) Tg(Zp3-cre) 93Knw; Zp3Cre de Vries et al., 2000 de Vries et al., 2000 de Vries et al., 2000 PMID:10686600 RRID:MGI :3835503 mixed background Strain, strain background (M. musculus) 129-Alpl tm(cre)Nagy Lomelı́ et al., 2000 Lomelı́ et al., 2000 Lomelı́ et al., 2000 PMID:10686602 RRID:IMSR_ JAX:008569 mixed background Antibody mouse anti-CDX2 Biogenex BioGenex Cat# AM392; RRID:AB_2650531 (1:200) Antibody goat anti -SOX2 Neuromics Neuromics Cat# GT15098; RRID:AB_2195800 (1:200) Antibody rabbit anti-PARD6B Novus Biologicals Novus Cat# NBP1-87337; RRID:AB_11034389 (1:100) Antibody rabbit anti-PARD6B Santa Cruz Biotechnology Santa Cruz Biotechnology Cat# sc-67393; RRID:AB_2267889 (1:100) Antibody mouse anti-PKCz Santa Cruz Biotechnology Santa Cruz Biotechnology Cat# sc-17781; RRID:AB_628148 (1:100) Antibody mouse anti-YAP1 Santa Cruz Biotechnology Santa Cruz Biotechnology Cat# sc-101199; RRID:AB_1131430 (1:200) Antibody mouse anti-pYAP1 Cell Signaling Technology Cell Signaling Technology Cat# 4911; RRID:AB_2218913 (1:800) Antibody chicken anti-GFP Aves Labs Aves Labs Cat# GFP-1020; RRID:AB_10000240 (1:2000) Continued on next page Frum et al. eLife 2018;7:e42298.

Techniques: Expressing, Control, Membrane, Inhibition, Staining, Marker, Injection, Cell Culture, Immunofluorescence, Confocal Microscopy

Journal: eLife

Article Title: HIPPO signaling resolves embryonic cell fate conflicts during establishment of pluripotency in vivo

doi: 10.7554/elife.42298

Figure Lengend Snippet: Figure 3. LATS2 directs inner cell mass fate independently of Sox2 (A) Lats2 and GFP or GFP alone were overexpressed in embryos lacking maternal or maternal and zygotic Sox2. (B) Lats2/GFP-overexpressing cells (dotted line) contribute almost exclusively to the inner cell mass in the presence or absence of Sox2 (n = embryos). (C) Proportion of non-injected cells and cells injected with Lats2/GFP mRNAs contributing to inner cell mass in the indicated genetic backgrounds. No significant differences were observed based on embryo genotype, indicating that Sox2 is dispensable for inside positioning by Lats2-overexpression (P, chi-squared test; n = embryos). (D) Proportion of non-injected cells and cells injected with the indicated mRNAs contributing to trophectoderm in the indicated genetic backgrounds. No significant differences were observed based on embryo genotype (P, chi- squared test; n = embryos). DOI: https://doi.org/10.7554/eLife.42298.008

Article Snippet: Key resources table Reagent type (species) or resource Designation Source or reference Identifiers Additional information Strain, strain background (Mus musculus) CD-1 Charles River Laboratories RRID:IMSR _CRL:22 Strain, strain background (M. musculus) Sox2tm1.1Lan Smith et al. (2009) PMID:19666824 RRID:IMSR_ JAX:013093 mixed background, Sox2 null refers to recombined allele Strain, strain background (M. musculus) Wwtr1 conditional allele; Wwtr1tm1.1Eno; Wwtr1loxp Xin et al., 2013 Xin et al., 2013 PMID:23918388 MGI:5544289 mixed background, ‘Wwtr1-’ or ‘Wwtr1 D ’ refers to recombined allele Strain, strain background (M. musculus) Yap1 conditional allele; Yap1tm1.1Eno; Yap1loxp Xin et al., 2011 Xin et al., 2011 PMID:22028467 MGI:5446483 mixed background, , ‘Yap1-’ or ‘Yap1 D ’ refers to recombined allele Strain, strain background (M. musculus) Tg(Zp3-cre) 93Knw; Zp3Cre de Vries et al., 2000 de Vries et al., 2000 de Vries et al., 2000 PMID:10686600 RRID:MGI :3835503 mixed background Strain, strain background (M. musculus) 129-Alpl tm(cre)Nagy Lomelı́ et al., 2000 Lomelı́ et al., 2000 Lomelı́ et al., 2000 PMID:10686602 RRID:IMSR_ JAX:008569 mixed background Antibody mouse anti-CDX2 Biogenex BioGenex Cat# AM392; RRID:AB_2650531 (1:200) Antibody goat anti -SOX2 Neuromics Neuromics Cat# GT15098; RRID:AB_2195800 (1:200) Antibody rabbit anti-PARD6B Novus Biologicals Novus Cat# NBP1-87337; RRID:AB_11034389 (1:100) Antibody rabbit anti-PARD6B Santa Cruz Biotechnology Santa Cruz Biotechnology Cat# sc-67393; RRID:AB_2267889 (1:100) Antibody mouse anti-PKCz Santa Cruz Biotechnology Santa Cruz Biotechnology Cat# sc-17781; RRID:AB_628148 (1:100) Antibody mouse anti-YAP1 Santa Cruz Biotechnology Santa Cruz Biotechnology Cat# sc-101199; RRID:AB_1131430 (1:200) Antibody mouse anti-pYAP1 Cell Signaling Technology Cell Signaling Technology Cat# 4911; RRID:AB_2218913 (1:800) Antibody chicken anti-GFP Aves Labs Aves Labs Cat# GFP-1020; RRID:AB_10000240 (1:2000) Continued on next page Frum et al. eLife 2018;7:e42298.

Techniques: Injection, Over Expression

Journal: eLife

Article Title: HIPPO signaling resolves embryonic cell fate conflicts during establishment of pluripotency in vivo

doi: 10.7554/elife.42298

Figure Lengend Snippet: Figure 5. Wwtr1 and Yap1 are required to repress SOX2 expression in outside cells. (A) CDX2 and SOX2 in wild type embryos at E3.25 (16–32 cell stages). CDX2 staining is more intense in outside cells than inside cells and SOX2 staining is specific to inside cells (n = embryos). (B) Embryos lacking maternal Wwtr1 and Yap1 with and heterozygous for Wwtr1 and Yap1 (which we consider to have 2 doses of WWTR1/YAP1) exhibit normal CDX2 and SOX2 expression (n = embryos). (C) Embryos lacking maternal Wwtr1 and Yap1 and heterozygous for either Wwtr1 or Yap1 (1 dose of WWTR1/YAP1) exhibit a high degree of ectopic SOX2 in outside cells (arrowheads), but continue to express CDX2, although the levels appear reduced (n = embryos). (D) Embryos lacking maternal and zygotic Wwtr1 and Yap1 (0 doses of WWTR1/YAP1) have the most severe phenotype, with a high degree of ectopic SOX2 in outside cells (arrowheads) and little or no detectable CDX2 (n = embryos). (E) Quantification of the percentage of outside cells in which ectopic SOX2 is detected in the presence of decreasing dose of Wwtr1 and Yap1 (t = student’s t-test, n = embryos). (F) Quantification of the percentage of outside cells in which CDX2 is detected in the presence of decreasing dose of Wwtr1 and Yap1 (t = student’s t-test, n = embryos). DOI: https://doi.org/10.7554/eLife.42298.010

Article Snippet: Key resources table Reagent type (species) or resource Designation Source or reference Identifiers Additional information Strain, strain background (Mus musculus) CD-1 Charles River Laboratories RRID:IMSR _CRL:22 Strain, strain background (M. musculus) Sox2tm1.1Lan Smith et al. (2009) PMID:19666824 RRID:IMSR_ JAX:013093 mixed background, Sox2 null refers to recombined allele Strain, strain background (M. musculus) Wwtr1 conditional allele; Wwtr1tm1.1Eno; Wwtr1loxp Xin et al., 2013 Xin et al., 2013 PMID:23918388 MGI:5544289 mixed background, ‘Wwtr1-’ or ‘Wwtr1 D ’ refers to recombined allele Strain, strain background (M. musculus) Yap1 conditional allele; Yap1tm1.1Eno; Yap1loxp Xin et al., 2011 Xin et al., 2011 PMID:22028467 MGI:5446483 mixed background, , ‘Yap1-’ or ‘Yap1 D ’ refers to recombined allele Strain, strain background (M. musculus) Tg(Zp3-cre) 93Knw; Zp3Cre de Vries et al., 2000 de Vries et al., 2000 de Vries et al., 2000 PMID:10686600 RRID:MGI :3835503 mixed background Strain, strain background (M. musculus) 129-Alpl tm(cre)Nagy Lomelı́ et al., 2000 Lomelı́ et al., 2000 Lomelı́ et al., 2000 PMID:10686602 RRID:IMSR_ JAX:008569 mixed background Antibody mouse anti-CDX2 Biogenex BioGenex Cat# AM392; RRID:AB_2650531 (1:200) Antibody goat anti -SOX2 Neuromics Neuromics Cat# GT15098; RRID:AB_2195800 (1:200) Antibody rabbit anti-PARD6B Novus Biologicals Novus Cat# NBP1-87337; RRID:AB_11034389 (1:100) Antibody rabbit anti-PARD6B Santa Cruz Biotechnology Santa Cruz Biotechnology Cat# sc-67393; RRID:AB_2267889 (1:100) Antibody mouse anti-PKCz Santa Cruz Biotechnology Santa Cruz Biotechnology Cat# sc-17781; RRID:AB_628148 (1:100) Antibody mouse anti-YAP1 Santa Cruz Biotechnology Santa Cruz Biotechnology Cat# sc-101199; RRID:AB_1131430 (1:200) Antibody mouse anti-pYAP1 Cell Signaling Technology Cell Signaling Technology Cat# 4911; RRID:AB_2218913 (1:800) Antibody chicken anti-GFP Aves Labs Aves Labs Cat# GFP-1020; RRID:AB_10000240 (1:2000) Continued on next page Frum et al. eLife 2018;7:e42298.

Techniques: Expressing, Staining

Journal: eLife

Article Title: HIPPO signaling resolves embryonic cell fate conflicts during establishment of pluripotency in vivo

doi: 10.7554/elife.42298

Figure Lengend Snippet: Figure 7. Resolution of cell fate conflicts in the preimplantation mouse embryo. (A) The expression of Sox2 and Cdx2 is differentially sensitive to YAP1/ WWTR1 activity, leading to co-expression of both lineage markers in cells when YAP1/WWTR1 activity levels are intermediate. (B) During division from the 16 to the 32-cell stage, cells that inherit the apical membrane repress HIPPO signaling and maintain an outside position. However, cells that inherit a smaller portion of the apical membrane would initially elevate their HIPPO signaling. We propose that elevated HIPPO then feeds back onto polarity by further antagonizing PAR-aPKC complex formation, leading to a snowball effect on repression of Sox2 expression, and thus ensuring that SOX2 is never detected in outside cells because these cells are rapidly internalized or apoptosed. (C) A closeup of the boxed region in panel B. In most outside cells, low LATS2 activity enables high levels of YAP1/WWTR1 activity, which repress Sox2 and apoptosis and promote Cdx2 expression and apical localization of aPKC and PARD6B, which in turn repress the HIPPO pathway. In rare outside cells, LATS2 activity becomes elevated, leading to lower activity of YAP1/WWTR1, which then leads these cells to become internalized or to undergo apoptosis. DOI: https://doi.org/10.7554/eLife.42298.013

Article Snippet: Key resources table Reagent type (species) or resource Designation Source or reference Identifiers Additional information Strain, strain background (Mus musculus) CD-1 Charles River Laboratories RRID:IMSR _CRL:22 Strain, strain background (M. musculus) Sox2tm1.1Lan Smith et al. (2009) PMID:19666824 RRID:IMSR_ JAX:013093 mixed background, Sox2 null refers to recombined allele Strain, strain background (M. musculus) Wwtr1 conditional allele; Wwtr1tm1.1Eno; Wwtr1loxp Xin et al., 2013 Xin et al., 2013 PMID:23918388 MGI:5544289 mixed background, ‘Wwtr1-’ or ‘Wwtr1 D ’ refers to recombined allele Strain, strain background (M. musculus) Yap1 conditional allele; Yap1tm1.1Eno; Yap1loxp Xin et al., 2011 Xin et al., 2011 PMID:22028467 MGI:5446483 mixed background, , ‘Yap1-’ or ‘Yap1 D ’ refers to recombined allele Strain, strain background (M. musculus) Tg(Zp3-cre) 93Knw; Zp3Cre de Vries et al., 2000 de Vries et al., 2000 de Vries et al., 2000 PMID:10686600 RRID:MGI :3835503 mixed background Strain, strain background (M. musculus) 129-Alpl tm(cre)Nagy Lomelı́ et al., 2000 Lomelı́ et al., 2000 Lomelı́ et al., 2000 PMID:10686602 RRID:IMSR_ JAX:008569 mixed background Antibody mouse anti-CDX2 Biogenex BioGenex Cat# AM392; RRID:AB_2650531 (1:200) Antibody goat anti -SOX2 Neuromics Neuromics Cat# GT15098; RRID:AB_2195800 (1:200) Antibody rabbit anti-PARD6B Novus Biologicals Novus Cat# NBP1-87337; RRID:AB_11034389 (1:100) Antibody rabbit anti-PARD6B Santa Cruz Biotechnology Santa Cruz Biotechnology Cat# sc-67393; RRID:AB_2267889 (1:100) Antibody mouse anti-PKCz Santa Cruz Biotechnology Santa Cruz Biotechnology Cat# sc-17781; RRID:AB_628148 (1:100) Antibody mouse anti-YAP1 Santa Cruz Biotechnology Santa Cruz Biotechnology Cat# sc-101199; RRID:AB_1131430 (1:200) Antibody mouse anti-pYAP1 Cell Signaling Technology Cell Signaling Technology Cat# 4911; RRID:AB_2218913 (1:800) Antibody chicken anti-GFP Aves Labs Aves Labs Cat# GFP-1020; RRID:AB_10000240 (1:2000) Continued on next page Frum et al. eLife 2018;7:e42298.

Techniques: Expressing, Activity Assay, Membrane