Journal: Neuron

Article Title: Nicotinamide pathway dependent Sirt1 activation restores calcium homeostasis to achieve neuroprotection in spinocerebellar ataxia type 7

doi: 10.1016/j.neuron.2019.11.019

Figure Lengend Snippet: (A) Here we see a diagram of the cell membrane, membrane-based cell signaling pathways, and the ER, indicating proteins involved in regulating calcium homeostasis and flux. Genes that encode these proteins or regulators are shown in italic font. Red stars demarcate the 11 calcium regulatory genes that contain PPAR response elements (PPREs) in their promoters. (B) We prepared cerebellar protein lysates from 30 week-old fxSCA7-92Q mice and WT controls, and then performed immunoprecipitation of PGC-1α. Immunoblotting of PGC-1α IP’s with antibodies directed against either anti-acetyl-lysine or anti-PGC-1α was performed, revealing a marked increase in acetylated PGC-1α in SCA7 cerebellum. We quantified these results by densitometry analysis of the acetyl-lysine and PGC-1α bands in the graph below. WT: n=4, fxSCA7 92Q: n=4; two-tailed t-test, *P <0.05. (C) PARP1 immunoblot analysis of cerebellar protein lysates from 8.5 week-old SCA7 266Q mice and WT controls, quantified by densitometry analysis of PARP1 and β-actin (loading control) bands in the graph below. WT: n=4, SCA7 266Q: n=5; two-tailed t-test, *P <0.05. (D) Mass spectrometry measurement of NAD+ levels in the cerebellum and cortex of 8.5 week-old SCA7-266Q mice and WT controls. WT: n=5, SCA7 266Q: n=5; two-tailed t-test, *P <0.05. (E) γH2Ax immunostaining of cortical neurons cultured from SCA7 210Q k.i. mice and WT littermates, at baseline and after 1 hr of H202. LEFT: Representative image set. γH2Ax (red), DAPI (blue). RIGHT: Quantification of γH2Ax+ signal / soma area; n = 3 mice/genotype; n = 3 cultures / mouse; n = 50 neurons / culture; ***P <.001, *P <.05, two-tailed t-test. Scale bar = 20 μm. Error bars = s.e.m.

Article Snippet: Anti-PGC-1α (H-300) Antibody (Rabbit Polyclonal Antibody) , Santa Cruz Biotechnology , Cat# sc-13067; RRID:AB_2166218.

Techniques: Membrane, Protein-Protein interactions, Immunoprecipitation, Western Blot, Two Tailed Test, Control, Mass Spectrometry, Immunostaining, Cell Culture

Journal: Neuron

Article Title: Nicotinamide pathway dependent Sirt1 activation restores calcium homeostasis to achieve neuroprotection in spinocerebellar ataxia type 7

doi: 10.1016/j.neuron.2019.11.019

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Anti-PGC-1α (H-300) Antibody (Rabbit Polyclonal Antibody) , Santa Cruz Biotechnology , Cat# sc-13067; RRID:AB_2166218.

Techniques: Produced, Virus, Plasmid Preparation, Recombinant, Protease Inhibitor, Western Blot, Reverse Transcription, cDNA Synthesis, SYBR Green Assay, Bicinchoninic Acid Protein Assay, Calcium Assay, Staining, shRNA, Software

Journal: Cell Genomics

Article Title: A highland-adaptation variant near MCUR1 reduces its transcription and attenuates erythrogenesis in Tibetans

doi: 10.1016/j.xgen.2025.100782

Figure Lengend Snippet:

Article Snippet: Mouse monoclonal anti-PGC1α , Proteintech , Cat#66369-1-lg; RRID: AB_2828002.

Techniques: Recombinant, Modification, Protease Inhibitor, Luminescence Assay, DNA Extraction, Multiplex Assay, Reporter Assay, Reverse Transcription, SYBR Green Assay, Electrophoresis, Mobility Shift, Transfection, Chromatin Immunoprecipitation, Mutagenesis, Enzyme-linked Immunosorbent Assay, Purification, Sequencing, RNA Sequencing, Gene Expression, Expressing, Real-time Polymerase Chain Reaction, Plasmid Preparation, Software

Journal: Molecular Medicine

Article Title: CAMKK2 restored mitochondrial dynamics homeostasis to alleviate pulmonary fibrosis via AMPK/PGC-1α signaling pathway in lung fibroblasts

doi: 10.1186/s10020-025-01373-5

Figure Lengend Snippet: The alterations of CAMKK2 expression and mitochondrial dynamics in fibrotic lungs and TGF-β1 stimulated fibroblasts. A-C Re-analysis of the CAMKK2 , PPARGC1A and MFN2 in the GSE47460 microarray datasets. D - E Representative images and quantification of IHC staining for CAMKK2 in lung sections from IPF patients and non-IPF control ( n = 5), Scale bars = 50 μm. F - G Representative images and quantification of IHC staining for CAMKK2 in lung sections from bleomycin induced mice and saline control ( n = 5), Scale bars = 50 μm. H - I Western blotting analysis and quantification of CAMKK2, COL1A1, PGC-1α, MFN2 and MFN1 levels in mouse lung homogenates ( n = 3). J - K Western blotting analysis and quantification of CAMKK2, α-SMA, PGC-1α, MFN2, MFN1 and DRP1 levels in MRC-5 cells treated for 48 h with and without TGF-β1 (5 ng/mL) ( n = 3). L Representative images of co-staining of CAMKK2 with α-SMA (a fibroblast marker) in lung tissues from bleomycin induced mice and saline control ( n = 5). Scale bars = 50 μm. The data represent one of at least three independent experiments. Data are shown as the mean ± SD. * P < 0.05; ** <0.01; *** P < 0.001; ns = not significant

Article Snippet: A selective CAMKK2 inhibitor STO-609(HY-19805), a potent AMPK inhibitor Compound C (dorsomorphin dihydrochloride, HY-13418 A), a specific PGC-1α inhibitor SR-18,292 (HY-101491) were obtained from MCE (Shanghai, China).

Techniques: Expressing, Microarray, Immunohistochemistry, Control, Saline, Western Blot, Staining, Marker

Journal: Molecular Medicine

Article Title: CAMKK2 restored mitochondrial dynamics homeostasis to alleviate pulmonary fibrosis via AMPK/PGC-1α signaling pathway in lung fibroblasts

doi: 10.1186/s10020-025-01373-5

Figure Lengend Snippet: CAMKK2 modulated mitochondrial dynamics homeotasis via the AMPK/PGC-1α signaling pathway. A - B Western blotting analysis and quantification of p T172 -AMPKα and t-AMPKα in MRC-5 cells transfected with pcDNA3.1 plasmid or pcDNA3.1-Flag-CAMKK2 plasmid for 24 h, then stimulated with TGF-β1 (5 ng/mL) for 24 h ( n = 3). C - D Western blotting analysis and quantification of p T172 -AMPKα, t-AMPKα, PGC-1α, MFN2 and MFN1 in MRC-5 cells transfected with pcDNA3.1 plasmid or pcDNA3.1-Flag-CAMKK2 plasmid for 24 h, followed with pre-treatment with Compound C (a potent AMPK inhibitor, 10µM) for 2 h, before stimulation with 5 ng/mL TGF-β1( n = 3). E - F Western blotting analysis and quantification of MFN2 and MFN1 in MRC-5 cells transfected with pcDNA3.1 plasmid or pcDNA3.1-Flag-CAMKK2 plasmid for 24 h, followed with pre-treatment with SR-18,292 (a specific PGC-1α inhibitor, 20µM) for 2 h, before stimulation with 5 ng/mL TGF-β1( n = 3), as below. G ROS relative level of MRC-5 cells ( n = 5). H ATP content (µmol/mg prot) of MRC-5 cells( n = 5). I - J The OCR (pmol/min) of MRC-5 cells ( n = 5). K - L Western blotting analysis and quantification of ND1-C-I, CYTB-C-III, MT-CO2-C-IV and ATP8-C-V in MRC-5 cells ( n = 3). M - N Western blotting analysis and quantification of FN1, COL1A1 and α-SMA in MRC-5 cells( n = 3). O - P Representative images and quantification of contraction activity by collagen gel contraction assay in MRC-5 cells ( n = 3). The data represent one of at least three independent experiments. Data are shown as the mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001. ns = not significant

Article Snippet: A selective CAMKK2 inhibitor STO-609(HY-19805), a potent AMPK inhibitor Compound C (dorsomorphin dihydrochloride, HY-13418 A), a specific PGC-1α inhibitor SR-18,292 (HY-101491) were obtained from MCE (Shanghai, China).

Techniques: Western Blot, Transfection, Plasmid Preparation, Activity Assay, Collagen Gel Contraction Assay

Journal: Molecular Medicine

Article Title: CAMKK2 restored mitochondrial dynamics homeostasis to alleviate pulmonary fibrosis via AMPK/PGC-1α signaling pathway in lung fibroblasts

doi: 10.1186/s10020-025-01373-5

Figure Lengend Snippet: Camkk2 attenuated bleomycin induced pulmonary fibrosis in mice. C57BL/6 N mice were treated with AAV6, AAV6-Camkk2-Flag, and bleomycin challenge in mice, as described in methods. A The schematic diagram depicts the timeline of the experiment. B Representative micro-CT images of mouse lungs on the day 14 after bleomycin challenge. C Representative images of H&E and Masson trichrome staining of lung sections. Scale bars = 100 μm. D Ashcroft score of mice( n = 6). E Lung/body weight of mice ( n = 6). F BALF total cell counts of mice ( n = 6). G Protein level in BALF of mice ( n = 6). H Hydroxyproline content in the left lung ( n = 6). I mRNA expression of Camkk2 , Fn1 , Col1a1 , and Acta2 in mouse lung homogenates assessed by qRT-PCR ( n = 6). J - L Western blotting analysis and quantification of Vimentin, α-SMA, PGC-1α, MFN2 and MFN1 levels in mouse lung homogenates ( n = 3). The data represent one of at least three independent experiments. Data are shown as the mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001. ns = not significant

Article Snippet: A selective CAMKK2 inhibitor STO-609(HY-19805), a potent AMPK inhibitor Compound C (dorsomorphin dihydrochloride, HY-13418 A), a specific PGC-1α inhibitor SR-18,292 (HY-101491) were obtained from MCE (Shanghai, China).

Techniques: Micro-CT, Staining, Expressing, Quantitative RT-PCR, Western Blot

Journal: Molecular Medicine

Article Title: CAMKK2 restored mitochondrial dynamics homeostasis to alleviate pulmonary fibrosis via AMPK/PGC-1α signaling pathway in lung fibroblasts

doi: 10.1186/s10020-025-01373-5

Figure Lengend Snippet: Schematic overview of mechanism (By Figdraw). The proposed model shows that CAMKK2 overexpression inhibits fibroblasts activation and myofibroblasts differentiation by reinstating mitochondrial dynamics via the AMPK/PGC-1α signaling pathway to alleviate pulmonary fibrosis

Article Snippet: A selective CAMKK2 inhibitor STO-609(HY-19805), a potent AMPK inhibitor Compound C (dorsomorphin dihydrochloride, HY-13418 A), a specific PGC-1α inhibitor SR-18,292 (HY-101491) were obtained from MCE (Shanghai, China).

Techniques: Over Expression, Activation Assay