phsl  (Cell Signaling Technology Inc)

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

Journal: bioRxiv

Article Title: Functionally Mature Bioengineered Human Skeletal Muscle Tissues Capture Essential Aspects of Glucose Metabolism

doi: 10.64898/2026.01.16.698404

Figure Lengend Snippet: (A) Effect of acute treatment with clenbuterol (10 nM, 20 min) on phosphorylation of β-adrenergic (B2AR) downstream targets including HSL (Ser660), CREB (Ser133), and RPS6 (Ser235/236) (n=4). (B) Gene expression of B2AR–responsive transcripts ( PPARGC1A , NR4A3 , NR4A1 , PGK1 , GYS1 , and others) after chronic clenbuterol treatment at the indicated doses (n=5). (C) Maximal tetanic force measured over time in tissues exposed to increasing clenbuterol concentrations (0.2–10 nM). Right: day-7 maximal force relative to vehicle for each dose (n=3). (D) Representative confocal images of myofibrillar structure in vehicle- and clenbuterol-treated tissues (n=2) (E) Maximal tetanic force during an inactivity protocol with or without clenbuterol treatment. Left: force traces over the detraining period. Right: day-7 maximal force in vehicle versus clenbuterol-treated groups (n=5). (F) Fatigue development during repeated contractions (protocol 2) in glucose-containing media in control versus clenbuterol-treated tissues. Inset: endurance values at the end of the protocol (n=5). (G) Fatigue development during repeated contractions (protocol 2) in glucose-free media in control versus clenbuterol-treated tissues (n=5). Data are presented as mean ± SEM, with individual values shown where applicable. Statistical comparisons were performed using one-way or two-way ANOVA with appropriate multiple-comparisons correction. ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: The following antibodies were used: SERCA2 (SCBT sc-53010), Hexokinase II (SCBT sc-130358), Glycogen synthase (was a kind gift from Oluf Pedersen, University of Copenhagen, DK), Phosphofructokinase (SCBT sc-166722), Mito cocktail (NDUFB8, SDHB, UQCRC2, COX-II, ATP5A) (Abcam ab110411), Citrate Synthase (Abcam ab96600), AMPK α 1 (a kind gift from Olga Göransson, Lund University, SE), α 2 (MRC PPU Reagents and Services, University of Dundee, Scotland, UK, custom made), β 1 (SCBT sc-100357), β 2 (1.5, a kind gift from Dr. DG Hardie, University of Dundee, Scotland, UK), γ 1 (Abcam ab32508), γ 3 (Genscript, NJ, USA, custom made), Myogenin (Abcam ab1835), MyoD1 (Thermo Scientific, Waltham, MA, PA5-23078), pHSL Ser660 (CST #4126), pCREB Ser133 (CST #9198), pRPS6 Ser 236/236 (CST #2211), pRPS6 Ser240/244 (CST #2215), GLUT4 (Thermo Scientific, Waltham, MA, PA1-1065), PDH-E1α (SC-377092).

Techniques: Phospho-proteomics, Gene Expression, Control