uncoupler  (MedChemExpress)

Journal: The Journal of Biological Chemistry

Article Title: CircAMOTL1 promotes adipose lipolysis and browning in cancer cachexia through miR-211-5p-mediated TET2 activation

doi: 10.1016/j.jbc.2025.111052

Figure Lengend Snippet: CircAMOTL1 promotes lipolysis and WAT browning in adipocytes. A and B , qPCR and Western blot analysis of the expression of adipose-related markers in adipocytes without/with overexpression of circAMOTL1. C , ORO staining of lipid accumulation in adipocytes without/with overexpression of circAMOTL1 (Scale bar: 100 μm). D and E , qPCR and Western blot analysis of the expression of adipose-related markers in adipocytes without/with knockdown of circAMOTL1. F , ORO staining of lipid accumulation in adipocytes without/with knockdown of circAMOTL1 (Scale bar: 100 μm). G , IF of differentiated adipocytes staining for mitochondria (MitoTracker, green ), anti-UCP1 ( red ), and nuclei (DAPI, blue ) (Scale bar: 100 μm).

Article Snippet: After blocking, membranes were incubated overnight at 4 °C with primary antibodies against TET2 (Abcam, ab124297), adipose triglyceride lipase (ATGL) (Abcam, ab207799), hormone-sensitive lipase (HSL) (Cell signaling technology, 4107), uncoupling protein 1 (UCP1) (Cell signaling technology, 72298), peroxisome proliferator-activated receptor-gamma coactivator (PGC)-1alpha (Abcam, ab191838), and GAPDH.

Techniques: Western Blot, Expressing, Over Expression, Staining, Knockdown

Journal: The Journal of Biological Chemistry

Article Title: CircAMOTL1 promotes adipose lipolysis and browning in cancer cachexia through miR-211-5p-mediated TET2 activation

doi: 10.1016/j.jbc.2025.111052

Figure Lengend Snippet: TET2 promotes lipolysis and WAT browning in adipocytes. A and B , qPCR and Western blot analysis of the expression of adipose-related markers in adipocytes without/with overexpression of TET2. C , ORO staining of lipid accumulation in adipocytes without/with overexpression of TET2 (Scale bar: 100 μm). D and E , qPCR and Western blot analysis of the expression of adipose-related markers in adipocytes without/with knockdown of TET2. F , ORO staining of lipid accumulation in adipocytes without/with knockdown of TET2 (Scale bar: 100 μm). G , IF of differentiated adipocytes staining for mitochondria (MitoTracker, green ), anti-UCP1 ( red ), and nuclei (DAPI, blue ) (Scale bar: 100 μm).

Article Snippet: After blocking, membranes were incubated overnight at 4 °C with primary antibodies against TET2 (Abcam, ab124297), adipose triglyceride lipase (ATGL) (Abcam, ab207799), hormone-sensitive lipase (HSL) (Cell signaling technology, 4107), uncoupling protein 1 (UCP1) (Cell signaling technology, 72298), peroxisome proliferator-activated receptor-gamma coactivator (PGC)-1alpha (Abcam, ab191838), and GAPDH.

Techniques: Western Blot, Expressing, Over Expression, Staining, Knockdown

Journal: The Journal of Biological Chemistry

Article Title: CircAMOTL1 promotes adipose lipolysis and browning in cancer cachexia through miR-211-5p-mediated TET2 activation

doi: 10.1016/j.jbc.2025.111052

Figure Lengend Snippet: CircAMOTL1 promotes lipolysis and WAT browning in vivo . A , schematic diagram, representative images, and H&E staining of subcutaneous adipose tissues in four different groups of mice. B , weight changes in four different groups of mice after injections. C , comparison of harvested subcutaneous WAT from the four experimental groups (from top to bottom , AAV-Vector, AAV-circAMOTL1, AAV-Vector + C26, and AAV-circAMOTL1 + C26). D , the expression of circAMOTL1 and protein levels of HSL, ATGL, UCP1, PGC1α and TET2 in adipose tissues from four different groups were analyzed.

Article Snippet: After blocking, membranes were incubated overnight at 4 °C with primary antibodies against TET2 (Abcam, ab124297), adipose triglyceride lipase (ATGL) (Abcam, ab207799), hormone-sensitive lipase (HSL) (Cell signaling technology, 4107), uncoupling protein 1 (UCP1) (Cell signaling technology, 72298), peroxisome proliferator-activated receptor-gamma coactivator (PGC)-1alpha (Abcam, ab191838), and GAPDH.

Techniques: In Vivo, Staining, Comparison, Plasmid Preparation, Expressing

Journal: Molecular Cancer Therapeutics

Article Title: The Recycling Collagen Receptor uPARAP Is a Unique Mediator of Stromal Drug Delivery to Carcinoma Cells

doi: 10.1158/1535-7163.MCT-25-0051

Figure Lengend Snippet: Targeting of uPARAP-negative tumors with a uPARAP-directed ADC. A and B, EBC-1 and HT29 carcinoma cells are insensitive to a uPARAP-directed ADC in vitro but sensitive to the free cytotoxin. A, EBC-1 cells (left), HT29 cells (middle), or uPARAP-expressing SaOS-2 cells (positive control; right) were cultured in vitro in the presence of the indicated concentrations of uPARAP-targeted ADC (9b7-MMAE) or nontargeted control ADC (aTNP-MMAE). After 72 hours of culture, the surviving cell fraction was determined by MTS assay. B, EBC-1 or HT29 cells were cultured in the presence of the indicated concentrations of uncoupled MMAE, after which the surviving cell fraction was determined as in A . C–E, Treatment of EBC-1 and HT29 tumors with 9b7-MMAE in vivo . EBC-1 or HT29 cells were injected into CB17 SCID mice as in Supplementary Figs. S2 and S5. Tumors were allowed to grow until reaching a size of 80 to 150 mm 3 , after which treatment was initiated with tail i.v. injections of the indicated doses of uPARAP-targeted 9b7-MMAE, nontargeted control ADC (aTNP-MMAE), unconjugated mAb 9b7 ( C only), or vehicle alone. Treatment was performed on days 0, 4, 7, and 11 (dotted lines). Tumor growth and regression were followed by palpation. For weight curves, see Supplementary Fig. S2. (Left) Tumor volume average ± SD for each treatment arm, ending the curve when the first mouse reached the humane endpoint. (Middle) Tumor volume in individual mice. (Right) Kaplan–Meier curves showing mouse survival for each treatment arm. ns, nonsignificant; P > 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001 by log-rank test with post hoc Bonferroni correction. C, n = 10 (PBS) and n = 9 (9b7 unconjugated, aTNP-MMAE, 9b7-MMAE). D and E, n = 8.

Article Snippet: Maleimide-derivatized monomethyl auristatin E (MMAE) linker-payload for coupling to IgG [maleimidocaproyl-valine-citrulline-p-aminobenzoyloxycarbonyl-MMAE (MC-VC-PAB-MMAE)] and uncoupled MMAE were commercial products from MedChemExpress (cat. no. HY-15575, CAS No. 646502-53-6) and Creative Biolabs (cat. no. ADC-P-022, CAS No. 474645-27-7), respectively.

Techniques: In Vitro, Expressing, Positive Control, Cell Culture, Control, MTS Assay, In Vivo, Injection