Journal: Cell metabolism

Article Title: Regulation of Tumor Initiation by the Mitochondrial Pyruvate Carrier

doi: 10.1016/j.cmet.2019.11.002

Figure Lengend Snippet: (A) Macroscopic tumor burden was assessed in Mpc1Lrig1KO, Mpc1Lrig1KO/+, and control mice at 100 days post-AOM (n=28–45 mice), ****p≤0.0001, ***p≤0.001, one-way ANOVA with multiple comparisons. Representative gross colon tumor burden and distribution are shown (ruler tick = 1mm). (B) At 100 days post-AOM, microscopic tumor grade and size assessed by pathologic grading in colons of Mpc1Lrig1KO, Mpc1Lrig1KO/+, and control mice (n=10–12), n.s.=not significant, *p≤0.05, one-way ANOVA with multiple comparisons on number of microscopic tumors per mouse. (C) Hematoxylin/Eosin and anti-β-catenin staining of control and Mpc1Lrig1KO adenomas at 100 days post-AOM, (100X, scale bars = 25μm). Black arrows indicate nuclear β-catenin. Green dashed circles indicate crypt necrosis. (D) Immunoblot of paired normal crypts (“C”) and tumor (“T”) samples from two control mice and two Mpc1Lrig1KO mice for MPC1, MPC2, and actin. Densitometry for MPC1 and MPC2, normalized to actin is shown with relative values adjusted to 100 for each control mouse normal crypt sample. (E) 100 days post-AOM, Ki67-positive nuclei as a percentage of total adenoma nuclei of Mpc1Lrig1KO and control adenomas were quantified on blinded high magnification images (n =5–7 mice, adenomas=10–20), **p≤0.01, unpaired two-tailed t-test. Representative images to the right (100X, scale bars=25μm). (F) 100 days post-AOM, cleaved caspase 3-positive nuclei of ApcLrig1KO/+Mpc1Lrig1KO and ApcLrig1KO/+ adenomas (black arrows) were quantified on blinded high magnification images (n =5–7 mice, adenomas=13–15, 100X, scale bars=25 μm), n.s.=not significant, unpaired two-tailed t-test. Green dashed circle indicates crypt necrosis and these positive nuclei were omitted. (G) At 49 days post-AOM, regions of superficial inflammation, epithelial injury with active inflammation (AI), aberrant foci, and low-grade dysplasia assessed by pathologic grading in colons of Mpc1Lrig1KO/+, Mpc1Lrig1KO/+, and control mice (n=8–10 mice), n.s.=not significant, unpaired t-test on number of microscopic lesions per mouse.

Article Snippet: b-catenin (IHC) 1:200 , BD Biosciences #610153 , RRID:AB_397554.

Techniques: Staining, Western Blot, Two Tailed Test

Journal: Cell metabolism

Article Title: Regulation of Tumor Initiation by the Mitochondrial Pyruvate Carrier

doi: 10.1016/j.cmet.2019.11.002

Figure Lengend Snippet: (A) Macroscopic tumor burden was assessed in Apc Lrig1KO/+Mpc1Lrig1KO, Apc Lrig1KO/+Mpc1Lrig1KO/+, and Apc Lrig1KO/+ mice at 90 days post-TAM (n=11–12 mice), **p≤0.01, *p≤0.05, one-way ANOVA with multiple comparisons (B) Microscopic tumor grade and size assessed by pathologic grading in the small intestine (left) and the colon (right) for ApcLrig1KO/+ Mpc1Lrig1KO and ApcLrig1KO/+ mice (n=5–7), n.s.=not significant, *p≤0.05, unpaired t-test on number of microscopic tumors per mouse. (C) Hematoxylin/Eosin, and anti-β-catenin staining of ApcLrig1KO/+ and ApcLrig1KO/+ Mpc1Lrig1KO adenomas (100X, scale bars=25 μm). Black arrows indicate nuclear β-catenin. (D) Immunoblot of paired normal crypts (“C”) and tumor (“T”) samples from two ApcLrig1KO/+ mice and two ApcLrig1KO/+Mpc1Lrig1KO mice for MPC1, MPC2, and actin. Densitometry for MPC1 and MPC2, normalized to actin is shown with relative values adjusted to 100 for each ApcLrig1KO/+ mouse normal crypt sample. (E) 90 days post-tamoxifen, Ki67-positive nuclei as a percentage of total adenoma nuclei of ApcLrig1KO/+Mpc1Lrig1KO and ApcLrig1KO/+ adenomas were quantified on blinded high magnification images (n =3–5 mice, adenomas=10–23), n.s.=not significant, unpaired two-tailed t-test. Representative images to the right (100X, scale bars=25 μm). (F) 90 days post-tamoxifen, cleaved caspase 3-positive nuclei of ApcLrig1KO/+Mpc1Lrig1KO and ApcLrig1KO/+ adenomas (black arrows) were quantified on blinded high magnification images (n =3–5 mice, adenomas = 15–16), n.s.= not significant, unpaired two-tailed t-test. Representative images to the right (100X, scale bars = 25 μm). Green dashed circle indicates crypt necrosis and these positive nuclei were omitted. (G) At 140 days post-TAM, macroscopic tumor burden per mouse in ApcVillKO/+ and ApcVillKO/+Mpc1VillKO was assessed (n=18–27,28: one small intestine was damaged during dissection and removed from analysis). *p≤0.05, unpaired t-test. (H) Microscopic tumor grade and size assessed by pathologic grading in the small intestine (left) and colon (right) of ApcVillKO/+ and ApcVillKO/+Mpc1VillKO mice (n=7–8), *p≤0.05, unpaired t-test on number of microscopic tumors per mouse.

Article Snippet: b-catenin (IHC) 1:200 , BD Biosciences #610153 , RRID:AB_397554.

Techniques: Staining, Western Blot, Two Tailed Test, Dissection

Journal: Cell metabolism

Article Title: Regulation of Tumor Initiation by the Mitochondrial Pyruvate Carrier

doi: 10.1016/j.cmet.2019.11.002

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: b-catenin (IHC) 1:200 , BD Biosciences #610153 , RRID:AB_397554.

Techniques: Western Blot, Recombinant, Plasmid Preparation, Protease Inhibitor, Generated, Software