Journal: Bone & Joint Research

Article Title: The promising regenerative potential of pulsed electromagnetic fields toward tendon differentiation

doi: 10.1302/2046-3758.152.BJR-2024-0550.R2

Figure Lengend Snippet: Scleraxis expression and fluorescence quantification in hamstring tendon explant sections with or without exposure to pulsed electromagnetic fields (PEMF) for 21 days, 8 hr/day. Representative images are shown. Left: representative immunostaining images for b) no PEMF and d) PEMF groups and their negative controls (a and c). Image d shows increased fluorescence intensity compared to image b. Yellow arrows in the images indicate representative localization of the specific markers. Scale bar: 25 μm. Right: graph depicting the results for each marker including the statistical analysis. Results are shown as mean (standard error of the mean) of corrected total cell fluorescence (CTCF). *Statistically significant p-value (< 0.05) in a two-tailed independent-samples t -test. Nine samples were evaluated.

Article Snippet: Immunohistochemistry stainings were performed using the Caspase-3 Cleaved (Asp175) antibody #9661 (Cell Signaling Technology, USA) with the avidin-biotin-peroxidase techniques (Anti-Mouse HRP-DAB Cell & Tissue Staining Kit, R&D Systems, USA).

Techniques: Expressing, Fluorescence, Immunostaining, Marker, Two Tailed Test

Journal: Bone & Joint Research

Article Title: The promising regenerative potential of pulsed electromagnetic fields toward tendon differentiation

doi: 10.1302/2046-3758.152.BJR-2024-0550.R2

Figure Lengend Snippet: c-Fos expression and fluorescence quantification in hamstring tendon explant sections with or without exposure to pulsed electromagnetic fields (PEMF) for 21 days, 8 hr/day. Representative images are shown. Left: representative immunostaining images for b) no PEMF and d) PEMF groups and their negative controls (a and c). Image d shows increased fluorescence intensity compared to image b. Yellow arrows in the images indicate representative localization of the specific markers. Scale bar: 25 μm. Right: graph depicting the results for each marker including the statistical analysis. Results are shown as mean (standard error of the mean) of corrected total cell fluorescence (CTCF). *Statistically significant p-value (< 0.05) in a two-tailed independent-samples t -test. Nine samples were evaluated.

Article Snippet: Immunohistochemistry stainings were performed using the Caspase-3 Cleaved (Asp175) antibody #9661 (Cell Signaling Technology, USA) with the avidin-biotin-peroxidase techniques (Anti-Mouse HRP-DAB Cell & Tissue Staining Kit, R&D Systems, USA).

Techniques: Expressing, Fluorescence, Immunostaining, Marker, Two Tailed Test

Journal: Bone & Joint Research

Article Title: The promising regenerative potential of pulsed electromagnetic fields toward tendon differentiation

doi: 10.1302/2046-3758.152.BJR-2024-0550.R2

Figure Lengend Snippet: Mammalian target of rapamycin (mTOR) expression and fluorescence quantification in hamstring tendon explant sections with or without exposure to pulsed electromagnetic fields (PEMF) for 21 days, 8 hr/day. Representative images are shown. Left: representative immunostaining images for b) no PEMF and d) PEMF groups and their negative controls (a and c). Image d shows increased fluorescence intensity compared to image b. Yellow arrows in the images indicate representative localization of the specific markers. Scale bar: 25 μm. Right: graph depicting the results for each marker including the statistical analysis. Results are shown as mean (standard error of the mean) of corrected total cell fluorescence (CTCF). *Statistically significant p-value (< 0.05) in a two-tailed independent-samples t -test. Nine samples were evaluated.

Article Snippet: Immunohistochemistry stainings were performed using the Caspase-3 Cleaved (Asp175) antibody #9661 (Cell Signaling Technology, USA) with the avidin-biotin-peroxidase techniques (Anti-Mouse HRP-DAB Cell & Tissue Staining Kit, R&D Systems, USA).

Techniques: Expressing, Fluorescence, Immunostaining, Marker, Two Tailed Test

Journal: Bone & Joint Research

Article Title: The promising regenerative potential of pulsed electromagnetic fields toward tendon differentiation

doi: 10.1302/2046-3758.152.BJR-2024-0550.R2

Figure Lengend Snippet: Proliferating cell nuclear antigen (PCNA) expression and fluorescence quantification in hamstring tendon explant sections with or without exposure to pulsed electromagnetic fields (PEMF) for 21 days, 8 hr/day. Representative images are shown. Left: representative immunostaining images for b) no PEMF and d) PEMF groups and their negative controls (a and c). No significant difference was observed in the number of PCNA-positive cells between the two groups. Scale bar: 25 μm. Right: graph depicting the results for each marker including the statistical analysis. Results are shown as mean (standard error of the mean) of the ratio between the number of PCNA-positive cells and the total number of cells in each field. Nine samples were evaluated. ns, non-significant.

Article Snippet: Immunohistochemistry stainings were performed using the Caspase-3 Cleaved (Asp175) antibody #9661 (Cell Signaling Technology, USA) with the avidin-biotin-peroxidase techniques (Anti-Mouse HRP-DAB Cell & Tissue Staining Kit, R&D Systems, USA).

Techniques: Expressing, Fluorescence, Immunostaining, Marker

Journal: Bone & Joint Research

Article Title: The promising regenerative potential of pulsed electromagnetic fields toward tendon differentiation

doi: 10.1302/2046-3758.152.BJR-2024-0550.R2

Figure Lengend Snippet: β-catenin expression and fluorescence quantification in hamstring tendon explant sections with or without exposure to pulsed electromagnetic fields (PEMF) for 21 days, 8 hr/day. Representative images are shown. Left: representative immunostaining images for b) no PEMF and d) PEMF groups and their negative controls (a and c). No significant difference was observed between the two groups. Image d is representative of the tendency toward decreased β-catenin signal. Yellow arrows in the images indicate representative localization of the specific markers. Scale bar: 25 μm. Right: graph depicting the results for each marker including the statistical analysis. Results are shown as mean (standard error of the mean) of corrected total cell fluorescence (CTCF). Nine samples were evaluated. ns, non-significant.

Article Snippet: Immunohistochemistry stainings were performed using the Caspase-3 Cleaved (Asp175) antibody #9661 (Cell Signaling Technology, USA) with the avidin-biotin-peroxidase techniques (Anti-Mouse HRP-DAB Cell & Tissue Staining Kit, R&D Systems, USA).

Techniques: Expressing, Fluorescence, Immunostaining, Marker

Journal: Bone & Joint Research

Article Title: The promising regenerative potential of pulsed electromagnetic fields toward tendon differentiation

doi: 10.1302/2046-3758.152.BJR-2024-0550.R2

Figure Lengend Snippet: Relative messenger RNA (mRNA) levels of a) scleraxis, b) collagen type I, c) collagen type VI, d) c-Fos, e) mammalian target of rapamycin (mTOR), f) proliferating cell nuclear antigen (PCNA), and g) β-catenin in tendon explants exposed to pulsed electromagnetic fields (PEMF) for 21 days, 8 hr/day, compared to tendon explants not exposed to PEMF measured by quantitative reverse transcription polymerase chain reaction analysis. Results were calculated as fold changes (mean (SD) of triplicates) relative to controls, normalized on 18S RNA level. *Statistically significant p-value (p < 0.05) in a two-tailed independent-samples t -test; ***Statistically significant p-value (p < 0.001) in a two-tailed independent-samples t -test. Three samples were evaluated. ns, non-significant.

Article Snippet: Immunohistochemistry stainings were performed using the Caspase-3 Cleaved (Asp175) antibody #9661 (Cell Signaling Technology, USA) with the avidin-biotin-peroxidase techniques (Anti-Mouse HRP-DAB Cell & Tissue Staining Kit, R&D Systems, USA).

Techniques: Reverse Transcription, Polymerase Chain Reaction, Two Tailed Test

Journal: eLife

Article Title: Amplification of a broad transcriptional program by a common factor triggers the meiotic cell cycle in mice

doi: 10.7554/eLife.43738

Figure Lengend Snippet:

Article Snippet: Commercial assay or kit , Mouse on Mouse ImmPRESS HRP (Peroxidase) Polymer Kit , VECTOR Laboratories , VectorLabs:MP2400 , .

Techniques: Knock-In, CRISPR, Generated, Recombinant, Plasmid Preparation, Sequencing, Software, Sample Prep, Blocking Assay, Fluorescence, Protease Inhibitor, Immunoprecipitation, Western Blot, Lysis

Journal: Molecular medicine reports

Article Title: HO‑1 alleviates cholesterol‑induced oxidative stress through activation of Nrf2/ERK and inhibition of PI3K/AKT pathways in endothelial cells.

doi: 10.3892/mmr.2017.6962

Figure Lengend Snippet: Figure 4. Measurements of ΔΨm, [Ca2+]i and [ROS]i in EA.hy926 cells stimulated with cholesterol. (A) Changes in the concentrations of [ROS]i in EA.hy926 cells following stimulation with cholesterol using confocal microscopy (magnification, x200). (B) The quantification results of mean fluorescence intensity of [ROS]i in EA.hy926 cells following stimulation with cholesterol. (C) Changes in the concentrations of [Ca2+]i in EA.hy926 cells following stimulation with cholesterol using confocal microscopy (magnification, x200). (D) The quantification results of mean fluorescence intensity of [Ca2+]i in EA.hy926 cells following stimulation with cholesterol. (E) Changes of ΔΨm in EA.hy926 cells following stimulation with cholesterol using confocal microscopy (magnifica tion, x200). (F) Ratio of JC‑1 polymer/JC‑1 monomer in EA.hy926 cells following stimulation with cholesterol. *P<0.05 vs. control group; #P<0.05 vs. SnPP group. HO‑1, heme oxygenase 1; SnPP, tin protoporphyrin; ΔΨm, mitochondrial membrane potential; [Ca2+]i, intercellular Ca2+; ROS, reactive oxygen species.

Article Snippet: The mitochondrial membrane potential (ΔΨm) assay kit with JC-1 (cat. no. C2006) was from Beyotime Institute of Biotechnology (Haimen, China).

Techniques: Confocal Microscopy, Fluorescence, Polymer, Control, Membrane

Journal: European Journal of Medical Research

Article Title: Shikonin induces the apoptosis and pyroptosis of EGFR -T790M-mutant drug-resistant non-small cell lung cancer cells via the degradation of cyclooxygenase-2

doi: 10.1186/s40001-024-02187-7

Figure Lengend Snippet: Shikonin induced the apoptosis and pyroptosis of H1975 cells. A MTT assay of H1975 cell viability after a 24 h treatment with 2 μM shikonin alone or in combination with a 3 h pre-treatment with PCD inhibitors, Z-VAD, CQ, Nec-1, and Fer-1. B Cell apoptosis analysis of H1975 cells treated with 0.5, 1, or 2 μM shikonin for 24 h. The apoptotic cells were quantified based on the data from three independent experiments. C The ΔΨm analysis of H1975 cells treated with 0.5, 1, or 2 μM shikonin for 24 h. The ΔΨm was calculated as the ratio of the JC-1 polymer (red) to the monomer (green). D Representative fluorescence images of the morphological features of H1975 cells treated with 0.5, 1, or 2 μM shikonin for 24 h. The white arrowheads indicate typical pyroptotic cells, characterized by PI + staining, swelling, and plasma membrane bubbling. PI + cells in five random fields were counted and statistically analyzed. Scale bar = 50 µm. E LDH release assay of H1975 cells treated with 0.5, 1, or 2 μM shikonin for 24 h. F , G WB analysis of H1975 cells treated with 0.5, 1, or 2 μM shikonin for 24 h or 2 μM shikonin for 6, 12, or 24 h. * p < 0.05, ** p < 0.01, *** p < 0.001, and ns: not significant

Article Snippet: The radioimmunoprecipitation assay (RIPA) buffer (P0013), phenylmethylsulfonyl fluoride (PMSF, ST506), N-acetyl-L-cysteine (NAC, S0077), annexin V-FITC cell apoptosis detection kit (C1062L), mitochondrial membrane potential detection kit (C2008S), lactate dehydrogenase (LDH) kit (C0016), chemiluminescence (ECL) kits (P0018S and P0018FS), and active oxygen detection kit (S0033S) were obtained from Beyotime (Shanghai, China).

Techniques: MTT Assay, Polymer, Fluorescence, Staining, Clinical Proteomics, Membrane, Lactate Dehydrogenase Assay

Journal: European Journal of Medical Research

Article Title: Shikonin induces the apoptosis and pyroptosis of EGFR -T790M-mutant drug-resistant non-small cell lung cancer cells via the degradation of cyclooxygenase-2

doi: 10.1186/s40001-024-02187-7

Figure Lengend Snippet: Shikonin induced the apoptosis and pyroptosis of H1975 cells by activating caspase-3. A MTT assay of H1975 cell viability after a 24 h treatment with 2 μM shikonin alone (shikonin-treated cells) or in combination with a 3 h pre-treatment with 25 μM DEVD (shikonin plus DEVD-treated cells). B Cell apoptosis analysis of the shikonin-treated and shikonin plus DEVD-treated H1975 cells. C The ΔΨm analysis of the shikonin-treated and shikonin plus DEVD-treated H1975 cells. The ΔΨm was calculated as the ratio of the JC-1 polymer (red) to the monomer (green). D , E Representative fluorescence images of the morphological features of the shikonin-treated and shikonin plus DEVD-treated H1975 cells. The white arrowheads indicate typical pyroptotic cells, characterized by PI + staining, swelling, and plasma membrane bubbling. PI + cells in five random fields were counted and statistically analyzed. Scale bar = 50 µm. F LDH release assay of the shikonin-treated and shikonin plus DEVD-treated H1975 cells. G WB analysis of the shikonin-treated and shikonin plus DEVD-treated H1975 cells. *** p < 0.001

Article Snippet: The radioimmunoprecipitation assay (RIPA) buffer (P0013), phenylmethylsulfonyl fluoride (PMSF, ST506), N-acetyl-L-cysteine (NAC, S0077), annexin V-FITC cell apoptosis detection kit (C1062L), mitochondrial membrane potential detection kit (C2008S), lactate dehydrogenase (LDH) kit (C0016), chemiluminescence (ECL) kits (P0018S and P0018FS), and active oxygen detection kit (S0033S) were obtained from Beyotime (Shanghai, China).

Techniques: MTT Assay, Polymer, Fluorescence, Staining, Clinical Proteomics, Membrane, Lactate Dehydrogenase Assay

Journal: European Journal of Medical Research

Article Title: Shikonin induces the apoptosis and pyroptosis of EGFR -T790M-mutant drug-resistant non-small cell lung cancer cells via the degradation of cyclooxygenase-2

doi: 10.1186/s40001-024-02187-7

Figure Lengend Snippet: Shikonin induced the apoptosis and pyroptosis of H1975 cells by increasing ROS production. A ROS levels of H1975 cells treated with 0.5, 1, or 2 μM shikonin for 24 h. B ROS levels of H1975 cells after a 24 h treatment with 2 μM shikonin alone (shikonin-treated cells) or in combination with a 3 h pre-treatment with 10 mM NAC (shikonin plus NAC-treated cells). C MTT assay of the shikonin-treated and shikonin plus NAC-treated H1975 cells. D Cell apoptosis analysis of the shikonin-treated and shikonin plus NAC-treated H1975 cells. E The ΔΨm analysis of the shikonin-treated and shikonin plus NAC-treated H1975 cells. The ΔΨm was calculated as the ratio of the JC-1 polymer (red) to the monomer (green). F Representative fluorescence images of the morphological features of the shikonin-treated and shikonin plus NAC-treated H1975 cells. The white arrowheads indicate typical pyroptotic cells, characterized by PI + staining, swelling, and plasma membrane bubbling. PI + cells in five random fields were counted and statistically analyzed. Scale bar = 50 µm. G LDH release assay of the shikonin-treated and shikonin plus NAC-treated H1975 cells. (H) WB analysis of the shikonin-treated and shikonin plus NAC-treated H1975 cells. ** p < 0.01, *** p < 0.001, and ns: not significant

Article Snippet: The radioimmunoprecipitation assay (RIPA) buffer (P0013), phenylmethylsulfonyl fluoride (PMSF, ST506), N-acetyl-L-cysteine (NAC, S0077), annexin V-FITC cell apoptosis detection kit (C1062L), mitochondrial membrane potential detection kit (C2008S), lactate dehydrogenase (LDH) kit (C0016), chemiluminescence (ECL) kits (P0018S and P0018FS), and active oxygen detection kit (S0033S) were obtained from Beyotime (Shanghai, China).

Techniques: MTT Assay, Polymer, Fluorescence, Staining, Clinical Proteomics, Membrane, Lactate Dehydrogenase Assay

Journal: European Journal of Medical Research

Article Title: Shikonin induces the apoptosis and pyroptosis of EGFR -T790M-mutant drug-resistant non-small cell lung cancer cells via the degradation of cyclooxygenase-2

doi: 10.1186/s40001-024-02187-7

Figure Lengend Snippet: COX-2 alleviated shikonin-induced apoptosis and pyroptosis of H1975 cells. A WB analysis of COX-2 expression in H1975/NC and H1975/COX-2 cells treated with 2 μM shikonin for 24 h. B ELISA of PGE2 levels in H1975/NC and H1975/COX-2 cells treated with 2 μM shikonin for 24 h. C MTT assay of the viability of H1975/NC and H1975/COX-2 cells treated with indicated concentrations of shikonin for 24 h. D Cell apoptosis analysis of H1975/NC and H1975/COX-2 cells treated with 2 μM shikonin for 24 h. E Representative fluorescence images of the morphological features of H1975/NC and H1975/COX-2 cells treated with 2 μM shikonin for 24 h. The white arrowheads indicate typical pyroptotic cells characterized by PI + staining, swelling, and plasma membrane bubbling. PI + cells in five random fields were counted and statistically analyzed. Scale bar = 50 µm. F LDH release assay of H1975/NC and H1975/COX-2 cells treated with 2 μM shikonin for 24 h. G , H WB analysis of H1975/NC and H1975/COX-2 cells treated with 2 μM shikonin for 24 h. ** p < 0.01, *** p < 0.001

Article Snippet: The radioimmunoprecipitation assay (RIPA) buffer (P0013), phenylmethylsulfonyl fluoride (PMSF, ST506), N-acetyl-L-cysteine (NAC, S0077), annexin V-FITC cell apoptosis detection kit (C1062L), mitochondrial membrane potential detection kit (C2008S), lactate dehydrogenase (LDH) kit (C0016), chemiluminescence (ECL) kits (P0018S and P0018FS), and active oxygen detection kit (S0033S) were obtained from Beyotime (Shanghai, China).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, MTT Assay, Fluorescence, Staining, Clinical Proteomics, Membrane, Lactate Dehydrogenase Assay

Journal: European Journal of Medical Research

Article Title: Shikonin induces the apoptosis and pyroptosis of EGFR -T790M-mutant drug-resistant non-small cell lung cancer cells via the degradation of cyclooxygenase-2

doi: 10.1186/s40001-024-02187-7

Figure Lengend Snippet: Celecoxib augmented the cytotoxic effects of shikonin by promoting the apoptosis and pyroptosis of H1975 cells. A MTT assay of the viability of H1975 cells treated with 2 μM shikonin and 60 μM celecoxib for 24 h (shikonin plus celecoxib-treated cells). B , C Cell apoptosis analysis of the shikonin plus celecoxib-treated H1975 cells. D Representative fluorescence images of the morphological features of the shikonin plus celecoxib-treated H1975 cells. The white arrowheads indicate typical pyroptotic cells characterized by PI + staining, swelling, and plasma membrane bubbling. PI + cells in five random fields were counted and statistically analyzed. Scale bar = 50 µm. E LDH release assay of the shikonin plus celecoxib-treated H1975 cells. F , G WB analysis of the shikonin plus celecoxib-treated H1975 cells. *** p < 0.001

Article Snippet: The radioimmunoprecipitation assay (RIPA) buffer (P0013), phenylmethylsulfonyl fluoride (PMSF, ST506), N-acetyl-L-cysteine (NAC, S0077), annexin V-FITC cell apoptosis detection kit (C1062L), mitochondrial membrane potential detection kit (C2008S), lactate dehydrogenase (LDH) kit (C0016), chemiluminescence (ECL) kits (P0018S and P0018FS), and active oxygen detection kit (S0033S) were obtained from Beyotime (Shanghai, China).

Techniques: MTT Assay, Fluorescence, Staining, Clinical Proteomics, Membrane, Lactate Dehydrogenase Assay

Journal: European Journal of Medical Research

Article Title: Shikonin induces the apoptosis and pyroptosis of EGFR -T790M-mutant drug-resistant non-small cell lung cancer cells via the degradation of cyclooxygenase-2

doi: 10.1186/s40001-024-02187-7

Figure Lengend Snippet: Schematic representation of the underlying molecular mechanisms of shikonin-induced apoptosis and pyroptosis of H1975 cells. Shikonin regulates COX-2 protein levels by inducing proteasomal degradation and ROS production, thereby inducing apoptosis and pyroptosis in H1975 cells

Article Snippet: The radioimmunoprecipitation assay (RIPA) buffer (P0013), phenylmethylsulfonyl fluoride (PMSF, ST506), N-acetyl-L-cysteine (NAC, S0077), annexin V-FITC cell apoptosis detection kit (C1062L), mitochondrial membrane potential detection kit (C2008S), lactate dehydrogenase (LDH) kit (C0016), chemiluminescence (ECL) kits (P0018S and P0018FS), and active oxygen detection kit (S0033S) were obtained from Beyotime (Shanghai, China).

Techniques: