Journal: Advanced Science

Article Title: H4K12 Lactylation Impairs Dopaminergic Neuron Energy Metabolism via the HIF‐1α/NDUFS1 Pathway in Parkinson's Disease

doi: 10.1002/advs.78220

Figure Lengend Snippet: H4K12la regulates HIF‐1α expression in PD models, and inhibition of lactylation by 2‐DG ameliorates related pathologies. (A) Western blotting analysis of H4K12la and HIF‐1α protein levels in SN4741 cells treated with 0.5 mM MPP + , NaLa, Lac, or NaCl for 24 h (N = 3). (B) Representative immunofluorescence images of SN4741 cells showing H4K12la (green) and HIF‐1α (red) expression in the nucleus under the indicated treatments. DAPI (blue) stains nuclei. Scale bar, 100 µm. Insets show 2 × magnifications of the boxed regions. (C) Quantitative analysis of the relative fluorescence intensity of H4K12la and HIF‐1α. (D,E) Western blotting analysis and quantification of H4K12la and HIF‐1α protein levels in the (D) striatum and (E) substantia nigra of MPTP‐induced PD mice (n = 3). (F, G) RT‐qPCR analysis of HIF‐1α mRNA levels in the (F) striatum and (G) substantia nigra of MPTP‐induced PD mice (n = 3). (H,I) Cells transfected with H4‐WT or H4‐K12R were treated with MPP + . H4K12la and HIF‐1α protein levels were assessed by Western blotting (N = 3). (J) Intracellular lactate levels in MPP + ‐treated SN4741 cells with or without 2‐DG treatment (N = 3). (K) Western blotting analysis of H4K12la and HIF‐1α protein levels in MPP + ‐treated SN4741 cells with or without 2‐DG co‐treatment (N = 3). (L) Representative immunofluorescence images of SN4741 cells showing that 2‐DG co‐treatment reduces MPP + ‐treated elevations of H4K12la (green) and HIF‐1α (red). Scale bar, 100 µm. (M) Corresponding quantification of fluorescence intensity for H4K12la and HIF‐1α. (N,O) Lactate levels in the (N) substantia nigra and (O) striatum homogenates of MPTP‐induced PD mice with or without 2‐DG treatment (n = 3). (P,Q) Western blotting analysis and quantification of H4K12la and HIF‐1α protein levels in the (P) striatum and (Q) substantia nigra of MPTP‐induced PD mice with or without 2‐DG treatment (n = 3). (R) Representative immunofluorescence images of midbrain sections from MPTP‐induced PD mice with or without 2‐DG treatment, showing TH (red) and H4K12la (green) expression. Scale bar, 200 µm (n = 3). Data are presented as mean ± SEM. ns, not significant, * p < 0.05, ** p < 0.01, and *** p < 0.001. Statistical significance was determined by one‐way ANOVA or Student's t‐test as appropriate. Lac, lactate; NaLa, sodium lactate; NaCl, sodium chloride; 2‐DG, 2‐deoxy‐D‐glucose. “n” refers to the number of mice per group. “N” refers to the number of biologically independent experiments.

Article Snippet: Mice in the 2‐DG + MPTP group were pretreated with 2‐DG (400 mg/kg, MCE, #HY‐13966) via intraperitoneal injection once daily for 3 days prior to MPTP administration.

Techniques: Expressing, Inhibition, Western Blot, Analysis, Immunofluorescence, Fluorescence, Quantitative RT-PCR, Transfection

Journal: Advanced Science

Article Title: H4K12 Lactylation Impairs Dopaminergic Neuron Energy Metabolism via the HIF‐1α/NDUFS1 Pathway in Parkinson's Disease

doi: 10.1002/advs.78220

Figure Lengend Snippet: 2‐DG ameliorates energy metabolism deficits, provides neuroprotection, and improves motor function in PD models by regulating H4K12la. (A,B) Measurement of (A) the ATP levels and (B) NAD + /NADH ratio in SN4741 cells treated with MPP + , with or without 2‐DG (N = 3). (C) Experimental schedule. (D–G) Measurement of the ATP levels and NAD + /NADH ratio in the (D,E) striatum and (F,G) substantia nigra of MPTP‐induced mice treated with or without 2‐DG (n = 3). (H) Representative immunohistochemical images and quantitative analysis of TH expression in the midbrain of mice treated with MPTP, with or without 2‐DG (n = 3). Scale bar, 200 µm. (I,J) Western blotting analysis and quantification of ATPB protein levels in the (I) striatum and (J) substantia nigra of mice treated with MPTP, with or without 2‐DG (n = 3). (K) RT‐qPCR analysis of TH mRNA levels in the substantia nigra of mice treated with MPTP, with or without 2‐DG (n = 3). (L–N) Behavioral assessments of MPTP‐induced mice with or without 2‐DG treatment: (L) time in the pole climbing test. (M) open‐field test showing representative images of mouse movement trajectories and total distance traveled. (N) Scores in the grasping test. n = 7 per group. Data are presented as mean ± SEM. * p < 0.05, ** p < 0.01, and *** p < 0.001. Statistical significance was determined by one‐way ANOVA or Student's t‐test as appropriate. ATPB, ATP synthase subunit beta. “n” refers to the number of mice per group. “N” refers to the number of biologically independent experiments.

Article Snippet: Mice in the 2‐DG + MPTP group were pretreated with 2‐DG (400 mg/kg, MCE, #HY‐13966) via intraperitoneal injection once daily for 3 days prior to MPTP administration.

Techniques: Immunohistochemical staining, Analysis, Expressing, Western Blot, Quantitative RT-PCR

Journal: Journal of Sport and Health Science

Article Title: Exercise attenuates stress-related signaling as sensed by higher phosphorylation of small heat shock proteins in skeletal muscle from older individuals

doi: 10.1016/j.jshs.2025.101111

Figure Lengend Snippet: Relative HSP abundances in whole skeletal muscle homogenates from young adults and older adults pre and post HIT exercise. Representative Westen blots of (A) HSP72, HSP27, and αB-crystallin and (B) phosphorylated HSP27 Ser15 (pHSP27 Ser15) and pαB-crystallin Ser59 in whole muscle homogenates from the vastus lateralis of the same individuals. Calibration curves of mixed muscle homogenates are indicated and were used to determine the relative number of given proteins (see Methods). Stain-free gels are indicative of total protein loading, and molecular weights are indicated by markers collected under white light capture without moving the membrane between that and chemiluminescence detection. Relative abundances of (C) HSP72, (D) HSP27, (E) pHSP27 Ser15, (F) αB-crystallin, and (G) pαB-crystallin Ser59 from young (circle) and older adults Pre (square) and older adults Post (triangle) HIT exercise are shown relative to average Old (pre) on a given gel (data are presented as mean ± SD). Individuals indicated by the number of symbols ( n : 5–7), with the same color assigned to the same individual and consistent across all graphs. * p ≤ 0.05 indicates Brown-Forsye and Welch’s and post hoc analysis using Games-Horwell. HIT = high-intensity training; HSP = heat shock protein; pαB-crystallin Ser59 = phospho-αB-crystallin at Serine59; pHSP27 Ser15 = phospho-HSP27 at Serine15.

Article Snippet: Details of antibodies used are as follows: HSP72 (1 in 500 mouse monoclonal, SMC100A; StressMarq Biosciences, Victoria, Canada); HSP27 (1 in 1000 mouse monoclonal, G3.1 ab2790; Abcam, Cambridge, UK); pHSP27 Ser15 (1 in 2000 monoclonal rabbit, ab76313; Abcam), pHSP27 Ser82 (1 in 2000 polyclonal mouse, ADI-SPA-524; Enzo Biochem, Farmingdale, NY, USA), αB-crystallin (1 in 1000 mouse monoclonal, SPA-222; StressGen Biotechnologies), pαB-crystallin Ser59 (1 in 1000 rabbit polyclonal, SPA-227; StressGen Biotechnologies).

Techniques: Staining, Membrane

Journal: Journal of Sport and Health Science

Article Title: Exercise attenuates stress-related signaling as sensed by higher phosphorylation of small heat shock proteins in skeletal muscle from older individuals

doi: 10.1016/j.jshs.2025.101111

Figure Lengend Snippet: HSP abundances in type I and II skeletal muscle fibers from young and older adults. (A, C, and F) The MHC isoform present was determined in individual muscle fiber segments from the vastus lateralis and, following pooling into type I and type II groups from a given biopsy, were analyzed by Westen blotting. Westen blots of (A) HSP72, (C) HSP27 and pHSP27 Ser15, (F) αB-crystallin and pαB-crystallin Ser59, with MHC isoforms in groups of fibers. Stain-free gels are indicative of total protein loading, and molecular weights are indicated by markers collected under white light capture without moving the membrane between that and chemiluminescence detection. Calibration curves of mixed muscle homogenates are indicated. Relative protein abundances of (B) HSP72, (D) HSP27, (E) pHSP27 Ser15, (G) αB-crystallin, and (H) pαB-crystallin Ser59 in fibers from young (circle) and older adults (square) type I fibers (no outline) and type II fibers (outline). All fibers are expressed relative to the average older adult’s type I fibers. The same color is assigned to the same individual and is consistent with (data are presented as mean ± SD). * p < 0.05 and ** p < 0.01, mixed effect model Univariant using either Tukey’s or Games-Horwell’s multiple comparison test (see Methods). HIT = high-intensity training; HSP = heat shock protein; MHC = myosin heavy chain; pαB-crystallin Ser59 = phospho-αB-crystallin at Serine59; pHSP27 Ser15 = phospho-HSP27 at Serine15.

Article Snippet: Details of antibodies used are as follows: HSP72 (1 in 500 mouse monoclonal, SMC100A; StressMarq Biosciences, Victoria, Canada); HSP27 (1 in 1000 mouse monoclonal, G3.1 ab2790; Abcam, Cambridge, UK); pHSP27 Ser15 (1 in 2000 monoclonal rabbit, ab76313; Abcam), pHSP27 Ser82 (1 in 2000 polyclonal mouse, ADI-SPA-524; Enzo Biochem, Farmingdale, NY, USA), αB-crystallin (1 in 1000 mouse monoclonal, SPA-222; StressGen Biotechnologies), pαB-crystallin Ser59 (1 in 1000 rabbit polyclonal, SPA-227; StressGen Biotechnologies).

Techniques: Staining, Membrane, Comparison

Journal: Journal of Sport and Health Science

Article Title: Exercise attenuates stress-related signaling as sensed by higher phosphorylation of small heat shock proteins in skeletal muscle from older individuals

doi: 10.1016/j.jshs.2025.101111

Figure Lengend Snippet: HSP abundances in type I and II skeletal fibers from older adults pre- and post HIT exercise. Relative protein abundances of (A and B) HSP72, (C and D) HSP27, (E and F) pHSP27 Ser15, (G and H) αB-crystallin, and (I and J) pαB-crystallin Ser59 in fibers from old pre and old post HIT exercise. All fibers are expressed relative to the average old pre type I fibers or relative pre type II depending on fiber type. The same color is assigned to the same Individual, consistent in both graphs and all figures. * p < 0.05 and ** p < 0.01 indicated significant difference in paired t -test (except pHSP27 Ser15 Wilcoxon match-pair rank test). Representative blots are shown in . HIT = high-intensity training; HSP = heat shock protein; pαB-crystallin Ser59 = phospho-αB-crystallin at Serine59; pHSP27 Ser15 = phospho-HSP27 at Serine15.

Article Snippet: Details of antibodies used are as follows: HSP72 (1 in 500 mouse monoclonal, SMC100A; StressMarq Biosciences, Victoria, Canada); HSP27 (1 in 1000 mouse monoclonal, G3.1 ab2790; Abcam, Cambridge, UK); pHSP27 Ser15 (1 in 2000 monoclonal rabbit, ab76313; Abcam), pHSP27 Ser82 (1 in 2000 polyclonal mouse, ADI-SPA-524; Enzo Biochem, Farmingdale, NY, USA), αB-crystallin (1 in 1000 mouse monoclonal, SPA-222; StressGen Biotechnologies), pαB-crystallin Ser59 (1 in 1000 rabbit polyclonal, SPA-227; StressGen Biotechnologies).

Techniques: