Journal: Neoplasia (New York, N.Y.)
Article Title: White-to-brown adipose switching promotes bladder cancer progression
doi: 10.1016/j.neo.2026.101282
Figure Lengend Snippet: PTHrP induces adipose browning through the PKA signaling pathway. (A-H) Adipocytes were subjected to two experimental treatments: (A, C, E, and G) adipocytes were treated with 100 ng/mL PTHrP or DMSO (vehicle control) in the presence or absence of H89 (50 µM), (B, D, F, and H) adipocytes with or without PTHR knockout were treated with bladder cancer cell-derived CM or control CM. (A and B) Representative immunoblots (left) of phosphorylated PKA substrates, p-HSL (Ser660), total HSL, p-CREB (Ser133), and total CREB in adipocytes with indicated treatments, with quantification (right) of the pHSL/total HSL and p-CREB/total CREB ratios. (C and D) Representative immunoblots (left) and quantification (right) of UCP1 in indicated adipocytes. (E and F) FFA release levels of indicated adipocytes were measured using a colorimetric assay. (G and H) Representative Oil Red O staining (left) and quantification (right) of neutral lipids in indicated adipocytes. Scale bars, 50 μm. Data were expressed as means ± SEM (A-H). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 by one-way ANOVA (A-H).
Article Snippet: The membranes were then incubated with primary antibodies including anti-UCP1 (1:3000, 83870-1-RR, Proteintech, China), anti-PTHLH (1:2000, A3183, ABclonal, China), anti-PTHR (1:1000, 29115-1-AP, Proteintech, China), anti-Phospho-PKA Substrate (RRXS*/T*) (1:1000, 9624, CST, USA), anti-pHSL (Ser660) (1:1000, AF8026, Affinity, China), anti-HSL (1:1000, AF6403, Affinity, China), anti-pCREB (Ser133) (1:2000, 28792-1-AP, Proteintech, China), anti-CREB(1:5000, 12208-1-AP, Proteintech, China), anti-GAPDH (1:50000, HRP-60004, Proteintech, China) and anti-ɑTubulin (1:10000, HRP-80762, Proteintech, China) overnight at 4°C.
Techniques: Control, Knock-Out, Derivative Assay, Western Blot, Colorimetric Assay, Staining
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