Journal: BMC Musculoskeletal Disorders

Article Title: The effect of Miya on skeletal muscle changes by regulating gut microbiota in rats with osteoarthritis through AMPK pathway

doi: 10.1186/s12891-024-08203-5

Figure Lengend Snippet: The histological changes of rats in the different groups observed by hematoxylin–eosin (HE) staining at 40 × , 100 × and 200 × magnification times. The green arrow indicates the surface of the cartilage; the yellow arrow points to cells; the red arrow indicates the tide line; and the blue arrow indicates the absence of cells. The rough triangle represents the nucleus; and the thin arrow represents the cytoplasm. Control: the rats without any treatments; OA: the osteoarthritis (OA) rat model; OA + MY: the OA rats treated with Miya (MY); OA + MY + adenosine 5’-monophosphate-activated protein kinase (AMPK) inhibitor: the OA rats firstly treated with MY, and then treated with BML275 (dorsomorphin, an AMPK inhibitor)

Article Snippet: After 2 weeks of modeling, the rats in the OA + MY, and OA + MY + AMPK inhibitor groups were firstly orally given MY (4 × 10 5 colony-forming units/mL C. butyricum , Miyalisan Pharmaceutical Co., Ltd, Japan) daily for 4 weeks, and then were intramuscularly injected with an equal volume of PBS, and 10 mg/kg BML275 (dorsomorphin, an AMPK inhibitor, cat. no. S31490, Shanghai Yuanye Biotechnology Co., Ltd, China) [ ] once a week for 4 weeks, respectively.

Techniques: Staining, Control

Journal: Adipocyte

Article Title: Zyflamend, a unique herbal blend, induces cell death and inhibits adipogenesis through the coordinated regulation of PKA and JNK

doi: 10.1080/21623945.2020.1803642

Figure Lengend Snippet: Inhibition of PKA and JNK abrogates the effects of Zyflamend on differentiation (a-b) Oil Red O-stained images of non-differentiated (Day 1) and differentiated adipocytes (Day 12) treated or non-treated with AMPK (BML275), JNK (SP600125), PKA (H89) or apoptosis (Z-VAD.fmk) inhibitors for the entire duration of differentiation. (b) Oil Red O stain was extracted, and its absorbance (520 nm) was quantitated. Bar graph represents data from at least six independent experiments, and data are expressed as mean + SEM. * p < 0.05, ** p < 0.01 indicate significant difference between non-treated and Zyflamend-treated cells. † p < 0.05, †† p < 0.01 indicate significant difference between cells treated with the indicated inhibitor together with Zyflamend and cells treated with Zyflamend only. (c) Phase contrast images of non-differentiated (Day 1) and differentiated adipocytes (Day 12) treated or non-treated with Zyflamend, SP600125, H89, or Z-VAD.fmk (alone or in combination with Zyflamend). Images in the far right panels are a zoomed capture of the boxed areas on Day 12 of differentiation. Scale bar: 100 μm. (d-g) Immunoblots of adipogenic markers in 3T3-MBX cells treated or non-treated with Zyflamend (200 μg/ml) with or without PKA inhibitor (H89; d-e) or JNK inhibitor (SP600125; f-g). Lysates were blotted for β-actin to control for loading. Representative Immunoblots from three independent experiments are shown. e-g) Bar graphs represent the indicated protein normalized to β-Actin as means + SEM. * p < 0.05, ** p < 0.01 indicate significant difference between day 1 and day 12 of differentiation for each treatment. † p < 0.05, †† p < 0.01 indicate significant difference between inhibitor-treated and non-treated cells

Article Snippet: AMPK inhibitor (BML275) was purchased from Santa Cruz Biotechnology (Santa Cruz, CA).

Techniques: Inhibition, Staining, Western Blot, Control

Journal: Nature

Article Title: Metabolic gatekeeper function of B-lymphoid transcription factors

doi: 10.1038/nature21076

Figure Lengend Snippet: a, b , Heatmap of metabolomics (n = 2 per group, each in triplicate; a ) and Western blots (n = 4; b ) of patient-derived leukemia samples. c , Glucose uptake and ATP levels during B→ myeloid reprogramming (n = 4). d , Fold change of pre-B ALL cells carrying GFP-tagged Cre following Lkb1 -deletion upon reprogramming (n = 6). e , Viability of pre-B ALL cells following deletion of Lkb1 or Ampkα2 (n = 3 independent experiments). f , Kaplan-Meier analysis (Mantel-Cox log-rank test) of recipient mice (n = 7 per group) injected with pre-B ALL cells following 4-OHT-induced deletion of Lkb1 or Ampkα2 (24 h). g , Patient-derived pre-B ALL cells treated with BML275 as indicated or in combination with prednisolone (n = 3), assessed by Combination Index (CI). Data, mean (± s.d), assessed by two-tailed t -test ( c ) or two-way ANOVA ( d, e ). For gel source data, see .

Article Snippet: BML275 (water-soluble) and Imatinib were obtained from Santa Cruz Biotechnology and LC Laboratories, respectively.

Techniques: Western Blot, Derivative Assay, Injection, Two Tailed Test

Journal: Nature

Article Title: Metabolic gatekeeper function of B-lymphoid transcription factors

doi: 10.1038/nature21076

Figure Lengend Snippet: a, Human leukemia and lymphoma cells (n = 4 biological replicates for B cell lymphoma and pre-B ALL; n = 3 biological replicates for CML; each in triplicate) were treated with BML275 (72 hr), and relative viability was assessed. b , Apoptosis was examined by Annexin V/7AAD staining in patient-derived pre-B ALL samples (n = 3) upon treatment with BML275 (10 μmol/L). c , Phospho-ACC-S 79 in patient-derived pre-B ALL samples (n = 5) following overnight treatment with control (−) or BML275 (+, 10 μmol/L) was assessed. d, Phosphorylation of S6-S 235/236 and Akt-S 473 in patient-derived pre-B ALL samples (n = 6) following overnight treatment with control (−) or BML275 (+, 10 μmol/L) was assessed. e, Levels of AMP (normalized to cell numbers) in patient-derived CML (CML5 and CML6) and Ph + ALL (ICN1 and PDX2) cells following 12 h treatment with control or BML275 (10 μmol/L). Plotted is log 2 -transformed average relative amounts (amount in cells treated with BML275/average amount of in control; 2 cases per group; each in triplicate), and data were median-centered. Median-centering was performed separately for CML5, CML6, ICN1 and PDX2 samples. f, Patient-derived CML (left) and pre-B ALL (middle) cells as well as MYC -driven B cell lymphoma cell lines (right) were treated with BML275 (10 μmol/L) or vehicle control for 6 hr. Glycolytic profiles (ECAR) and mitochondrial functions (OCR) were measured (normalized to total protein; n = 6; f ). Data shown as mean (± e.f.) and assessed by two-tailed t -test ( e, f ).

Article Snippet: BML275 (water-soluble) and Imatinib were obtained from Santa Cruz Biotechnology and LC Laboratories, respectively.

Techniques: Staining, Derivative Assay, Control, Phospho-proteomics, Transformation Assay, Two Tailed Test