Journal: Cell Stress & Chaperones

Article Title: Proteomic and phenotypic profiling of replicative-senescent human foreskin fibroblasts under brief heat shock

doi: 10.1016/j.cstres.2026.100174

Figure Lengend Snippet: Expression of several senescence-associated proteins under HS conditions at 41 °C and 45 °C for 30 min in aged HFF-1 cells. (a) Detection of protein expression levels of GORAB, AURKB, RhoC, α-N-catenin, RRM2, AURKA, and PLK1 under 41 °C and 45 °C HS. Protein expression was quantified as fold-change relative to untreated controls (set to 1), normalized to GAPDH. (b) GORAB. (c) AURKB. (d) RhoC. (e) α-N-catenin. (f) RRM2. (g) AURKA. (h) PLK1. Representative blots were cropped only to improve presentation; the corresponding full-length, uncropped blots with molecular-weight markers and the associated GAPDH loading control are provided in Supplementary Figure S1 (the first lane corresponds to an unrelated sample and was not analyzed). Data are representative of three independent experiments, with results presented as mean ± SD. ** P < 0.01, *** P < 0.001, **** P < 0.0001, compared with the control group.

Article Snippet: Membranes were blocked with 5% nonfat milk for 1 h at room temperature and incubated overnight at 4 °C with primary antibodies against GORAB (Proteintech, Wuhan, China; 17798–1-AP; 1:1000), AURKA (Cell Signaling Technology, Danvers, MA, USA; 14475T; 1:1000), AURKB (Cell Signaling Technology; 28711T; 1:1000), RhoC (Cell Signaling Technology; 3430T; 1:1000), RRM2 (Cell Signaling Technology; 65939T; 1:2000), PLK1 (Cell Signaling Technology; 4513T; 1:1000), α-N-catenin (Cell Signaling Technology; 2163T; 1:1000), and GAPDH (Abcam, Cambridge, UK; ab125247; 1:10,000) as the loading control.

Techniques: Expressing, Molecular Weight, Control

Journal: Experimental & Molecular Medicine

Article Title: Osteoblast-derived osteomodulin restrains osteoclastogenesis via ITGB8/RRM2-mediated reduction of mitochondrial respiration and mitochondrial ATP production

doi: 10.1038/s12276-026-01682-7

Figure Lengend Snippet: a Volcano plot of all transcripts. Red dots indicate >2-fold change upregulation ( P < 0.05) among the control group, while blue dots indicate >2-fold change downregulation ( P < 0.05) among the control group. b The heat map illustrates the 20 most significantly differentially expressed genes between the control group and OMD-treated group. Red indicates upregulation, while blue indicates downregulation ( n = 3). c After a 1-day induction, the mRNA levels of Rrm2 were measured in both the treatment group (100 ng/ml recombinant OMD protein) and the control group using qRT–PCR ( n = 3). d , e After a 3-day induction, the protein levels of RRM2 were assessed by WB in BMMs treated with 100 ng/ml recombinant OMD protein and in the control group ( d ), followed by quantitative analysis of the protein ( e ) ( n = 3). f Effects of osalmid on BMMs viability at 48 h ( n = 6). g , h After a 4-day induction, TRAP staining was conducted on BMMs exposed to various concentrations of osalmid ( g ) followed by a quantitative analysis of the nuclei counts in TRAP-positive multinuclear cells ( h ) ( n = 3). Scale bar, 100 μm. i After a 1-day induction, the mRNA levels of osteogenic genes in BMMs treated with different concentrations of osalmid were assessed using qRT–PCR ( n = 3). j , k After a 3-day induction, the protein levels of osteogenic genes in BMMs treated with varying concentrations of osalmid were analyzed using WB ( j ), followed by quantitative analysis of the proteins ( k ) ( n = 3). l After treating BMMs with either a control vector or Rrm2 -overexpressing adenovirus, different treatments were administered, and the mRNA levels of osteogenic genes were quantified using qRT–PCR ( n = 3). m , n After a 4-day induction, TRAP staining was conducted on BMMs exposed to different treatments, followed by a quantitative analysis of the nuclei counts in TRAP-positive multinuclear cells ( n = 3). Scale bar, 100 μm. Data represent mean ± s.e.m. Experimental data for each quantitative analysis were replicated at least three times. Statistical significance was assessed using unpaired t -test ( c and e ) or one-way ANOVA ( f , h , i , k , l and n ).

Article Snippet: Adenovirus-mediated Rrm2 was purchased from GENECHEM.

Techniques: Control, Recombinant, Quantitative RT-PCR, Staining, Plasmid Preparation

Journal: Experimental & Molecular Medicine

Article Title: Osteoblast-derived osteomodulin restrains osteoclastogenesis via ITGB8/RRM2-mediated reduction of mitochondrial respiration and mitochondrial ATP production

doi: 10.1038/s12276-026-01682-7

Figure Lengend Snippet: a Relative abundance of mitochondria determined by qPCR of mt-Co2Ⅱ DNA normalized to β-globin ( n = 3). b , c After a 3-day induction, the protein levels of mitochondrial OXPHOS complexes were assessed by WB in BMMs treated with 20 μm osalmid and DMSO ( b ), followed by quantitative analysis of the proteins ( c ) ( n = 3). d Relative abundance of mitochondria determined by qPCR of mt-Co2Ⅱ DNA normalized to β-globin ( n = 3). e , f After treating BMMs with either a control vector or Rrm2 -overexpressing adenovirus, different treatments were applied, and 3 days post-osteoclastogenesis induction, the protein levels of mitochondrial OXPHOS complexes were assessed by WB ( e ), followed by quantitative analysis of the protein ( f ) ( n = 3). g , h After a 3-day induction, OCR was measured ( g ), including detailed parameters such as basal respiration, maximal respiration, ATP-linked respiration, spare respiratory capacity, nonmitochondrial respiration and proton leak ( h ). i , j After a 3-day induction, ECAR was measured ( i ), including detailed parameters such as glycolysis, glycolytic capacity and glycolytic reserve ( j ). k After a 3-day induction, the ATP production rate was measured ( n = 3). Data represent mean ± s.e.m. Experimental data for each quantitative analysis were replicated at least three times. Statistical significance was assessed using unpaired t -tests ( a and c ) or one-way ANOVA ( d , f , h , j and k ).

Article Snippet: Adenovirus-mediated Rrm2 was purchased from GENECHEM.

Techniques: Control, Plasmid Preparation

Journal: Experimental & Molecular Medicine

Article Title: Osteoblast-derived osteomodulin restrains osteoclastogenesis via ITGB8/RRM2-mediated reduction of mitochondrial respiration and mitochondrial ATP production

doi: 10.1038/s12276-026-01682-7

Figure Lengend Snippet: a The MS of integrin β8 (ITGB8). b Predicted interactions between OMD and ITGB8 based on the GeneMANIA database. c The detailed interaction network between OMD and ITGB8. The key residues of OMD (in blue) and ITGB8 (in green) are displayed as sticks, with residue chain identifiers indicated. Dashed yellow lines represent hydrogen bonds, and dashed red lines indicate π–π interactions, with distances labeled. d Representative immunofluorescence images showing colocalization of OMD/ITGB8 in BMMs treated with 100 ng/ml OMD. Scale bar, 5 μm. e HEK-293T cells were transfected with Flag-OMD, HA-ITGB8 or Flag-OMD and HA-ITGB8. Flag immunoprecipitates were analyzed by immunoblotting as outlined. f , g After treating BMMs with either control siRNA or si- Itgb8 , different treatments were applied, and 3 days after osteoclastogenesis induction, the protein levels of osteoclastogenesis markers, RRM2 and ITGB8 were assessed by WB ( f ), followed by quantitative analysis of the proteins ( g ) ( n = 3). h , i After a 4-day induction, TRAP staining was conducted on BMMs exposed to different treatments ( h ), followed by a quantitative analysis of the nuclei counts in TRAP-positive multinuclear cells ( i ) ( n = 3). Scale bar, 100 μm. j , k The levels of total and phosphorylated proteins in multiple signaling pathways at indicated time points (0, 30, 60 and 120 min) following OMD treatment were assessed by WB ( j ), followed by quantitative analysis of the proteins ( k ) ( n = 3). l RhoA-GTP levels were measured by ELISA following OMD treatment ( n = 3). m The occupancy of TEAD at the Rrm2 promoter was assessed by ChIP ( n = 3). n BMMs were transfected with control siRNA or si- Itgb8 and subsequently subjected to the indicated treatments; total and phosphorylated YAP levels were analyzed by WB ( n = 3). o After a 4-day induction, TRAP staining was conducted on BMMs exposed to different treatments ( n = 3). Scale bar, 100 μm. Data represent mean ± s.e.m. Experimental data for each quantitative analysis were replicated at least three times. Statistical significance was assessed using unpaired t -tests ( k – m ) and one-way ANOVA ( g and i ).

Article Snippet: Adenovirus-mediated Rrm2 was purchased from GENECHEM.

Techniques: Residue, Labeling, Immunofluorescence, Transfection, Western Blot, Control, Staining, Protein-Protein interactions, Enzyme-linked Immunosorbent Assay