Journal: Clinical and Translational Radiation Oncology

Article Title: Differential regulation of radioadaptation by quercetin between human normal and cancer cells

doi: 10.1016/j.ctro.2025.101099

Figure Lengend Snippet: Quercetin restores NRF2 nuclear translocation in radioadapted MCF10A cells. Representative immunofluorescence images of NRF2 (green) and DAPI (blue) and quantification of the nuclear-to-cytoplasmic NRF2 fluorescence ratio in MCF10A cells 24 h after 5 Gy irradiation with or without prior LDRT and quercetin treatment (Scale bar: 10 μm). Data is shown as mean ± SEM. * p < 0.05, ** p < 0.01.

Article Snippet: Cells were incubated with rabbit anti-human NRF2 primary antibody (1:200; Proteintech, 16396–1-AP), and detection was performed using goat anti-rabbit Alexa Fluor 488 (1:500, Invitrogen, 11001).

Techniques: Translocation Assay, Immunofluorescence, Fluorescence, Irradiation

Journal: Translational Cancer Research

Article Title: OX40L and IL-2 combination strategy for gastric cancer immunotherapy

doi: 10.21037/tcr-2025-707

Figure Lengend Snippet: Immunohistochemical and flow cytometric analysis of gastric cancer tissue. (A) H&E histochemical staining of tumor-free area (left), tumor margin (middle), and gastric tumor site (right). (B) Representative IHC images for CD3 + , CD4 + , CD8 + , FOXP3, OX40, and OX40L in tumor-free area (left column), tumor margin (middle column), and gastric tumor site (right column) sections. (C) Immunofluorescence staining for the co-expression of CD3 + and OX40 in gastric cancer with anti-CD3 and anti-OX40 antibodies and DAPI for nuclear staining. (D) Flow cytometry analysis of OX40 expression in CD3 cells obtained from PBMC of gastric cancer patients (upper panel) and OX40 expression on CD3 on TILs in gastric cancer patients (low panel). Statistical analysis of all data were performed using one-way ANOVA. Data are shown as mean ± standard deviation. ns, nonsignificant; *, P<0.05; ***, P<0.001. ANOVA, analysis of variance; DAPI, 4',6-diamidino-2-phenylindole; H&E, hematoxylin & eosin; IHC, immunohistochemistry; OX40L, OX40 ligand; PBMC, peripheral blood mononuclear cells; TILs, tumor-infiltrating lymphocytes.

Article Snippet: Immunohistochemistry (IHC) analyses were performed using specific primary rabbit anti-human CD3 monoclonal antibody (Proteintech Group, Wuhan, China), mouse anti-human OX40 and rabbit anti-human OX40L monoclonal antibodies (CST, Danvers, MA, USA), and rabbit anti-human CD4 (Proteintech Group, Wuhan, China), mouse anti-human CD8 (Proteintech Group, Wuhan, China) antibody, rabbit anti-human FOXP3 monoclonal antibodies (CST, Danvers, MA, USA).

Techniques: Immunohistochemical staining, Staining, Immunofluorescence, Expressing, Flow Cytometry, Standard Deviation, Immunohistochemistry

Journal: Translational Cancer Research

Article Title: Prognostic chromatin remodeling signature stratifies survival outcomes in lung adenocarcinoma patients

doi: 10.21037/tcr-2025-1699

Figure Lengend Snippet: GJB3 knockdown impairs the oncogenic behavior of LUAD cells. (A,B) Efficient knockdown of GJB3 was confirmed by Western blot and qRT-PCR in A549 and H1299 cells. (C) The proliferative capacity of cells was significantly reduced after GJB3 silencing, as measured by CCK‑8 assay. (D,E) GJB3 knockdown markedly attenuated the migration and invasion capabilities of LUAD cells, shown by representative Transwell images (stained with crystal violet) at 200× magnification (D) and their quantification (E). All data are shown as mean ± SD (n=3). ****, P<0.0001. CCK-8, Cell Counting Kit-8; LUAD, lung adenocarcinoma; OD, optical density; qRT-PCR, quantitative reverse transcription polymerase chain reaction; SD, standard deviation.

Article Snippet: The membranes were blocked with 5% skim milk and then incubated overnight at 4 °C with rabbit anti-human GJB3 primary antibody (1:1,000, Proteintech, 12880-1-AP) and mouse anti-human GAPDH primary antibody (1:5,000, Proteintech, 60004-1-Ig).

Techniques: Knockdown, Western Blot, Quantitative RT-PCR, CCK-8 Assay, Migration, Staining, Cell Counting, Reverse Transcription, Polymerase Chain Reaction, Standard Deviation

Journal: iScience

Article Title: Agonism of PIEZO1 prevents aggravated periodontitis with traumatic occlusion via MAPK signaling pathway

doi: 10.1016/j.isci.2025.113688

Figure Lengend Snippet: PIEZO1 plays a role in the aggravation of periodontitis induced by traumatic occlusion (A) Representative picture of IHC staining and semi-quantitative analysis of PIEZO1. Scale bar: 100 μm. (B) Representative picture of IHC staining and semi-quantitative analysis of RUNX2. Scale bar: 100 μm. (C) Representative picture of IHC staining and semi-quantitative analysis of IL-6. Representative histological images from a single representative sample in the same group. Scale bar: 100 μm. Consecutive sections were subjected to IHC staining of PIEZO1 (A), IHC staining of RUNX2 (B), and IHC staining of IL-6 (C). PDL, periodontal ligament; D, dentin; Ab, alveolar bone. Statistical significance was tested with Kruskal-Wallis test followed by Dunn’s post hoc test. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001; ns, not significant ( p > 0.05). Error bars represent mean ± SD ( n = 6 biological replicates).

Article Snippet: After blocking, cells were incubated overnight at 4°C with a rabbit anti-human PIEZO1 primary antibody (1:200; Biorbyt, Cambridge, UK).

Techniques: Immunohistochemistry

Journal: iScience

Article Title: Agonism of PIEZO1 prevents aggravated periodontitis with traumatic occlusion via MAPK signaling pathway

doi: 10.1016/j.isci.2025.113688

Figure Lengend Snippet: Yoda1 activates the MAPK pathway by promoting the expression of PIEZO1 (A) Representative picture of IHF staining showing expression of PIEZO1 in the periodontal ligament. (B) Representative picture of IHC staining showing expression of RUNX2 in the periodontal ligament. (C) Representative picture of IHF staining showing expression of p-JNK in the periodontal ligament. (D) Representative picture of IHF staining showing expression of ERK and p-ERK in the periodontal ligament. Scale bar: 200 μm. Representative histological images from a single representative sample in the same group. Consecutive sections were subjected to IHF staining of PIEZO1 (A), IHC staining of RUNX2 (B), IHF staining of p-JNK (C), and IHF staining of ERK and p-ERK (D) ( n = 6 biological replicates).

Article Snippet: After blocking, cells were incubated overnight at 4°C with a rabbit anti-human PIEZO1 primary antibody (1:200; Biorbyt, Cambridge, UK).

Techniques: Expressing, Staining, Immunohistochemistry

Journal: iScience

Article Title: Agonism of PIEZO1 prevents aggravated periodontitis with traumatic occlusion via MAPK signaling pathway

doi: 10.1016/j.isci.2025.113688

Figure Lengend Snippet: MCS loading on hPDLFs (A) Schematic of MCS loading on hPDLFs with a magnitude of 4,000 μstrain and a frequency of 0.5 Hz. The expression of PIEZO1 (B) and RUNX2 (C) detected by RT-qPCR and quantitative analysis. Statistical significance was tested with Kruskal-Wallis test followed by Dunn’s post hoc test. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001; ns, not significant ( p > 0.05). Error bars represent mean ± SD ( n = 3 biological replicates).

Article Snippet: After blocking, cells were incubated overnight at 4°C with a rabbit anti-human PIEZO1 primary antibody (1:200; Biorbyt, Cambridge, UK).

Techniques: Expressing, Quantitative RT-PCR

Journal: iScience

Article Title: Agonism of PIEZO1 prevents aggravated periodontitis with traumatic occlusion via MAPK signaling pathway

doi: 10.1016/j.isci.2025.113688

Figure Lengend Snippet: The effect of MCS loading on the expression of PIEZO1, osteogenic factors, and inflammatory factors in both normal and inflammatory hPDLFs (A) Gene expression of PIEZO1 , RUNX2 , OPN , ALP , IL-6 , and TNF was assessed by RT-qPCR. (B) Protein expression of PIEZO1, RUNX2, OPN, and IL-6 was detected by WB, and the semi-quantitative analysis was performed using ImageJ. (C) Expression of PIEZO1 analyzed by ICF. Scale bar: 100 μm. (D) Expression of Ca 2+ in hPDLFs. Scale bar: 200 μm. (E) ALP staining of hPDLFs with images captured under an inverted microscope. Scale bar: 500 μm. (F) ARS staining of hPDLFs with images captured under an inverted microscope. Scale bar: 500 μm. Statistical significance was tested with Kruskal-Wallis test followed by Dunn’s post hoc test. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001; ns, not significant ( p > 0.05). Error bars represent mean ± SD ( n = 3 biological replicates).

Article Snippet: After blocking, cells were incubated overnight at 4°C with a rabbit anti-human PIEZO1 primary antibody (1:200; Biorbyt, Cambridge, UK).

Techniques: Expressing, Gene Expression, Quantitative RT-PCR, Staining, Inverted Microscopy

Journal: iScience

Article Title: Agonism of PIEZO1 prevents aggravated periodontitis with traumatic occlusion via MAPK signaling pathway

doi: 10.1016/j.isci.2025.113688

Figure Lengend Snippet: Yoda1 upregulated the expression of PIEZO1 and osteogenic-related factors and activated the MAPK signaling pathway (A) RT-qPCR detected the expression of PIEZO1 . (B) CCK-8 detected cell proliferation under different concentrations of Yoda1. (C) Fluo-4 AM calcium ion fluorescence staining. Scale bar: 100 μm. (D) RT-qPCR analysis of PIEZO1 , RUNX2 , and IL-6 expression in hPDLFs treated with LPS and mechanical compression stress (MCS), with or without Yoda1. (E) Western blot analysis of PIEZO1, RUNX2, IL-6, p-ERK, ERK, p-JNK, JNK, p-p38, and p38 in the same groups. Quantitative densitometric analysis was performed using ImageJ. Statistical significance was tested with Mann-Whitney U test. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001; ns, not significant ( p > 0.05). Error bars represent mean ± SD ( n = 3 biological replicates).

Article Snippet: After blocking, cells were incubated overnight at 4°C with a rabbit anti-human PIEZO1 primary antibody (1:200; Biorbyt, Cambridge, UK).

Techniques: Expressing, Quantitative RT-PCR, CCK-8 Assay, Fluorescence, Staining, Western Blot, MANN-WHITNEY

Journal: iScience

Article Title: Agonism of PIEZO1 prevents aggravated periodontitis with traumatic occlusion via MAPK signaling pathway

doi: 10.1016/j.isci.2025.113688

Figure Lengend Snippet: PIEZO1 deficiency inhibited the phosphorylation of MAPK signaling pathway, suppressing osteogenesis and promoting inflammation (A) RT-qPCR analysis of PIEZO1 , RUNX2 , and IL-6 expression in hPDLFs treated with LPS/MCS, Yoda1, and the MEK inhibitor trametinib, alone or in combination. (B) Western blot analysis of PIEZO1, RUNX2, IL-6, p-ERK, ERK, p-JNK, JNK, p-p38, p38, p-MEK, and MEK under the same treatment conditions. (C) ARS staining of hPDLFs with images captured under an inverted microscope. Scale bar: 500 μm. (D) ALP staining of hPDLFs with images captured under an inverted microscope. Scale bar: 500 μm. Quantitative analysis was conducted using ImageJ. Statistical significance was tested with Kruskal-Wallis test followed by Dunn’s post hoc test. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001; ns, not significant ( p > 0.05). Error bars represent mean ± SD ( n = 3 biological replicates).

Article Snippet: After blocking, cells were incubated overnight at 4°C with a rabbit anti-human PIEZO1 primary antibody (1:200; Biorbyt, Cambridge, UK).

Techniques: Phospho-proteomics, Quantitative RT-PCR, Expressing, Western Blot, Staining, Inverted Microscopy

Journal: iScience

Article Title: Agonism of PIEZO1 prevents aggravated periodontitis with traumatic occlusion via MAPK signaling pathway

doi: 10.1016/j.isci.2025.113688

Figure Lengend Snippet: Schematic diagram illustrated the mechanisms by which traumatic occlusion aggravated periodontitis via PIEZO1 and its association with the MAPK signaling pathway This figure was created by Figdraw.

Article Snippet: After blocking, cells were incubated overnight at 4°C with a rabbit anti-human PIEZO1 primary antibody (1:200; Biorbyt, Cambridge, UK).

Techniques: