Journal: Cancer Medicine

Article Title: Role of Ca 2+ ‐Dependent Epithelial‐Mesenchymal Transition in Malignant Progression of Colorectal Cancer: Special Focus on REG Iα/ EDNRB

doi: 10.1002/cam4.71754

Figure Lengend Snippet: REG Iα promotes cell migration, invasion, and EMT via EDNRB. (A) Cell migration was assessed by Transwell assay. (B) Matrigel‐coated Transwell assay assessed cell invasion. (C) Western blot analysis of EMT‐related markers, including the epithelial marker E‐Cadherin and mesenchymal markers N‐Cadherin and Vimentin. β‐actin was used as the loading control. Data are presented as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: CHOP (Abcam, ab317378), Cleaved‐Caspase 3 (Abcam, ab32042), Cleaved‐PARP (Cell Signaling Technology, 9541), E‐Cadherin (BOSTER, PB9561), N‐Cadherin (BOSTER, A01577‐3), and Vimentin (BOSTER, BM4029).

Techniques: Migration, Transwell Assay, Western Blot, Marker, Control

Journal: Cancer Medicine

Article Title: Role of Ca 2+ ‐Dependent Epithelial‐Mesenchymal Transition in Malignant Progression of Colorectal Cancer: Special Focus on REG Iα/ EDNRB

doi: 10.1002/cam4.71754

Figure Lengend Snippet: The REG Iα‐EDNRB axis promotes cell migration, invasion, and EMT via the Ca 2+ signaling pathway. (A) Transwell assay assessed cell migration. (B) Cell invasion was assessed by Matrigel‐coated Transwell assay. (C) EMT‐related protein levels (E‐Cadherin, N‐Cadherin, Vimentin) were detected by Western blot. β‐actin served as the loading control. Data are presented as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: CHOP (Abcam, ab317378), Cleaved‐Caspase 3 (Abcam, ab32042), Cleaved‐PARP (Cell Signaling Technology, 9541), E‐Cadherin (BOSTER, PB9561), N‐Cadherin (BOSTER, A01577‐3), and Vimentin (BOSTER, BM4029).

Techniques: Migration, Transwell Assay, Western Blot, Control

Journal: Cancer Medicine

Article Title: Role of Ca 2+ ‐Dependent Epithelial‐Mesenchymal Transition in Malignant Progression of Colorectal Cancer: Special Focus on REG Iα/ EDNRB

doi: 10.1002/cam4.71754

Figure Lengend Snippet: The REG Iα‐EDNRB‐Ca 2+ axis promotes tumor growth and EMT in vivo. (A) Representative images of the xenograft tumors and excised tumor tissues from the indicated groups. (B) Statistical analysis of the final tumor weights. (C) Tumor growth curves measuring tumor volume over time. (D) H&E staining showed pathological changes in tumor tissues. (E) TUNEL assay detected cell apoptosis in tumor tissues. (F) Cell proliferation in tumor tissues was shown by Ki67 IHC staining. (G) Expression of REG Iα (Immunofluorescence, upper row) and EDNRB (IHC, lower row) in tumor tissues. (H) Western blot analysis of REG Iα, EDNRB, and p‐CaMKII protein levels in tumor tissues. β‐actin was used as the loading control. (I) Western blot analysis of EMT‐related proteins (E‐Cadherin, N‐Cadherin, Vimentin) in tumor tissues. β‐actin was used as the loading control. Data are presented as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: CHOP (Abcam, ab317378), Cleaved‐Caspase 3 (Abcam, ab32042), Cleaved‐PARP (Cell Signaling Technology, 9541), E‐Cadherin (BOSTER, PB9561), N‐Cadherin (BOSTER, A01577‐3), and Vimentin (BOSTER, BM4029).

Techniques: In Vivo, Staining, TUNEL Assay, Immunohistochemistry, Expressing, Immunofluorescence, Western Blot, Control

Journal: Nature Metabolism

Article Title: N -acetylaspartate from fat cells regulates postprandial body temperature

doi: 10.1038/s42255-025-01334-6

Figure Lengend Snippet: a , Schematic of metabolite profiling of SVF-derived differentiated adipocytes generated from Aspa WT and Aspa KO mice. Glucose incorporation into metabolite pools was measured by incubating cells with U- 13 C glucose. Isotopolog distribution for indicated metabolites was measured after 24 h using IC-MS targeted profiling. b , Relative abundance of intracellular NAA (n = 3 biological replicates/group). Data represent mean ± s.e.m. **** P < 0.0001 by unpaired two-tailed Student’s t -test. c , Schematic showing contribution of 13 C 6 -labeled glucose tracing into early pyrimidines via aspartate (m + 2 and m + 3 isotopologs), and R5P and downstream pyrimidines (m + 5 isotopologs). G6P, glucose-6-phosphate; 6-PG, 6-phosphogluconate; R5P, ribose-5-phosphate; F6P, fructose-6-phosphate; 3PG, 3-phosphoglycerate. d , Fractional labeling of aspartate, early and later pyrimidines, and the pentose phosphate pathway metabolite, R5P. DHOA, dihydroorotate; OMP, orotidine 5’-monophosphate; UMP, uridine monophosphate. Isotopolog data are corrected for 13 C natural abundance (n = 3 replicates/group). Data are mean ± s.e.m. * P < 0.05, ** P < 0.01, *** P < 0.001 by unpaired two-tailed Student’s t -test. e , Immunoblot of Aspa WT and Aspa KO primary SVF cells ± differentiation, probed for ASPA, phospho-CAD (S1859), total CAD, phospho-S6K (T389), total S6K, and ADIPOQ. HSP90 served as the loading control. Representative of 3 independent experiments. f , Representative Oil-Red O (ORO) staining of lipid accumulation in differentiated Aspa WT and Aspa KO SVF adipocytes. Scale bar is 0.783 cm. Panels a and c created with BioRender.com .

Article Snippet: The following antibodies were used: anti-ASPA (1:2,500, ab154503, Abcam), anti-ADIPOQ (1:500, GTX112777, GeneTex), anti-FABP4 (1:1,000, GTX116036, GeneTex), anti-PPARG (1:1,000, 2443, Cell Signaling), anti-phospho-CAD S1859 (1:500, 12662, Cell Signaling), anti-CAD (1:500, 11933, Cell Signaling), anti-phospho-p70S6K T389 (1:1,000, 9205, Cell Signaling), anti-p70S6K (1:1,000, 2708, Cell Signaling), anti-DHOA dehydrogenase (1:1,000, 14877-I-AP, Proteintech), anti-HSP90 (1:1,000, 4877, Cell Signaling), anti-phospho-TSC2 (1:1,000, 3617, Cell Signaling), anti-TSC2 (1:1,000, 4308, Cell Signaling), anti-phospho-4EBP1 T37/46 (1:1,000, 2855, Cell Signaling), anti-4EBP1 (1:1,000, 9644, Cell Signaling), anti-phospho-AKT S473 (1:1,000, 4060, Cell Signaling), anti-AKT (pan) (1:1,000, 4691, Cell Signaling) and Total OXPHOS Rodent WB Antibody Cocktail (1:1,000, ab110413, Abcam).

Techniques: Derivative Assay, Generated, Two Tailed Test, Labeling, Western Blot, Control, Staining

Journal: Nature Metabolism

Article Title: N -acetylaspartate from fat cells regulates postprandial body temperature

doi: 10.1038/s42255-025-01334-6

Figure Lengend Snippet: a , Schematic of fast/re-feeding experiment. Body temperature measurements and plasma of Aspa WT and Aspa KO male mice were taken following 16 h fast and after 6 h of refeeding. Necropsy data from fasted and refed mice: (b) Body weight, (c) vWAT and (d) scWAT depot weights, shown as % of body weight. Fasted (n = 5 WT,5 KO mice/group), refed (n = 6 WT,9 KO mice/group). e , Body temperature measured by rectal probe. Fasted (n = 10 WT,12 KO mice/group) and refed (n = 10 WT,12 KO mice/group). For ΔT (n = 10 WT,12 KO mice/group), data are mean ± s.e.m., *P < 0.05 by unpaired two-tailed Student’s t-test. f , Plasma leptin levels. Fasted (n = 9 WT,7 KO mice/group), refed (n = 8 WT,6 KO mice/group). g , Immunoblot of scWAT from Aspa WT and Aspa KO mice following fasting and refeeding, probed for phospho-CAD (S1859), total CAD (tCAD), DHODH, phospho-S6K (T389), total S6K (tS6K), and ADIPOQ. HSP90 served as the loading control (n = 3 mice/group/condition). b-f Data represented as box-and-whisker plots using the Min-to-Max method in GraphPad Prism: box limits, 25 th to 75 th percentiles; center line, median; whiskers, minimum and maximum values. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 by ordinary two-way ANOVA followed by Fisher LSD tests. Panel a created with BioRender.com .

Article Snippet: The following antibodies were used: anti-ASPA (1:2,500, ab154503, Abcam), anti-ADIPOQ (1:500, GTX112777, GeneTex), anti-FABP4 (1:1,000, GTX116036, GeneTex), anti-PPARG (1:1,000, 2443, Cell Signaling), anti-phospho-CAD S1859 (1:500, 12662, Cell Signaling), anti-CAD (1:500, 11933, Cell Signaling), anti-phospho-p70S6K T389 (1:1,000, 9205, Cell Signaling), anti-p70S6K (1:1,000, 2708, Cell Signaling), anti-DHOA dehydrogenase (1:1,000, 14877-I-AP, Proteintech), anti-HSP90 (1:1,000, 4877, Cell Signaling), anti-phospho-TSC2 (1:1,000, 3617, Cell Signaling), anti-TSC2 (1:1,000, 4308, Cell Signaling), anti-phospho-4EBP1 T37/46 (1:1,000, 2855, Cell Signaling), anti-4EBP1 (1:1,000, 9644, Cell Signaling), anti-phospho-AKT S473 (1:1,000, 4060, Cell Signaling), anti-AKT (pan) (1:1,000, 4691, Cell Signaling) and Total OXPHOS Rodent WB Antibody Cocktail (1:1,000, ab110413, Abcam).

Techniques: Clinical Proteomics, Two Tailed Test, Western Blot, Control, Whisker Assay

Journal: Nature Metabolism

Article Title: N -acetylaspartate from fat cells regulates postprandial body temperature

doi: 10.1038/s42255-025-01334-6

Figure Lengend Snippet: a , Schematic of the gavage experiment. Body temperature measurements and plasma collection in WT mice were performed following a 16-h fast, 15–30 min following gavage and after 1 h of refeeding. b , Body temperature measurements in fasted ( n = 13 water, 13 NAA), post-gavage ( n = 13 water, 13 NAA) and refed mice ( n = 8 water, 8 NAA). *P < 0.05 by an ordinary two-way ANOVA followed by Fisher LSD tests. c , Plasma NAA levels of fasted ( n = 4 water, 4 NAA), post-gavage ( n = 9 water, 9 NAA) and refed mice ( n = 4 water, 4 NAA). d , Tissue NAA levels in the liver, scWAT and vWAT after gavage ( n = 4 mice per group per condition). e , Relative abundance of CarbAsp across the liver, scWAT and vWAT following gavage ( n = 4 mice per group per condition). f , Relative abundance of plasma CarbAsp ( n = 9 mice per group per condition), OMP ( n = 9 mice per group per condition) and UMP ( n = 4 mice per group per condition) following gavage. g , Immunoblot of scWAT from Aspa WT mice following water or NAA gavage, probed for ASPA, phospho-CAD (pCAD) S1859, total CAD (tCAD), phospho-AKT S473, total AKT (tAKT), phospho-TSC2 T1462, total TSC2 (tTSC2), phospho-4EBP1 T37/46 and total 4EBP1 (t4EBP1). HSP90 served as the loading control ( n = 3 mice per group per condition). h , Model for NAA’s mechanism of action in regulating de novo pyrimidine synthesis. NAA binds within the ATCase domain of CAD, promoting increased CAD activity. Carbamoyl-P, carbamoyl phosphate; N -carbamoyl-asp, N -carbamoyl aspartate; DHO, dihydroorotase; DHODH, DHOA dehydrogenase; PPi, pyrophosphate; PRPP, phosphoribosyl pyrophosphate; UMPs, UMP synthase; UDP, uridine 5′-diphosphate; UTP, uridine 5′-triphosphate; CTP, cytidine 5′-triphosphate; CDP, cytidine 5′-diphosphate; CMP, cytidine 5′-monophosphate; UCK, uridine cytidine kinase. In b – f , data are represented as box-and-whisker plots using the Min-to-Max method in GraphPad Prism: box limits, 25th to 75th percentiles; centre line, median; whiskers, minimum and maximum values. In c – f , data are shown as log 10 -transformed values. * P < 0.05, ** P < 0.01, **** P < 0.0001 by an unpaired two-tailed Student’s t -test. NS, no significance. Panels a and h created with BioRender.com .

Article Snippet: The following antibodies were used: anti-ASPA (1:2,500, ab154503, Abcam), anti-ADIPOQ (1:500, GTX112777, GeneTex), anti-FABP4 (1:1,000, GTX116036, GeneTex), anti-PPARG (1:1,000, 2443, Cell Signaling), anti-phospho-CAD S1859 (1:500, 12662, Cell Signaling), anti-CAD (1:500, 11933, Cell Signaling), anti-phospho-p70S6K T389 (1:1,000, 9205, Cell Signaling), anti-p70S6K (1:1,000, 2708, Cell Signaling), anti-DHOA dehydrogenase (1:1,000, 14877-I-AP, Proteintech), anti-HSP90 (1:1,000, 4877, Cell Signaling), anti-phospho-TSC2 (1:1,000, 3617, Cell Signaling), anti-TSC2 (1:1,000, 4308, Cell Signaling), anti-phospho-4EBP1 T37/46 (1:1,000, 2855, Cell Signaling), anti-4EBP1 (1:1,000, 9644, Cell Signaling), anti-phospho-AKT S473 (1:1,000, 4060, Cell Signaling), anti-AKT (pan) (1:1,000, 4691, Cell Signaling) and Total OXPHOS Rodent WB Antibody Cocktail (1:1,000, ab110413, Abcam).

Techniques: Clinical Proteomics, Western Blot, Control, Activity Assay, Whisker Assay, Transformation Assay, Two Tailed Test