Journal: Biomedicines

Article Title: REDD1 Affects Proliferation, Apoptosis, Migration, and Colony Formation via p-ERK and p-JNK Signaling in Lung Adenocarcinoma Cells Under Hypoxia

doi: 10.3390/biomedicines13122918

Figure Lengend Snippet: REDD1 is overexpressed in LUAD. ( A ) Immunohistochemistry staining images (200X) of REDD1 expression in four pairs of LUAD and normal tissues. ( B ) Western blot analyses of REDD1 protein level in samples of four matched LUAD (T), para-carcinoma tissues (P), and normal tissues (N); β-tubulin was used as the control. ( C ) Patients with high REDD1 expression exhibit a significantly reduced overall survival compared to those with low REDD1 expression. (Logrank p, p -value from log-rank test; HR, hazard ratio; p (HR), survival probability; n (high), number of high REDD1 expression; n (low), number of low REDD1 expression; TPM, transcripts per million). ( D , E ) Protein expression of REDD1 in A549 and H1299 cells at different durations of hypoxia. E, n = 4; F, n = 6. * p < 0.05; ** p < 0.01.

Article Snippet: Antibodies that recognize REDD1 (10638-1-AP), matrix metallopeptidase 1 (MMP1) (10371-2-AP), matrix metallopeptidase 9 (MMP9) (60600-1-Ig), and B-cell lymphoma-2-associated X (Bax) (50599-2-Ig) were purchased from Proteintech (Wuhan, China).

Techniques: Immunohistochemistry, Staining, Expressing, Western Blot, Control

Journal: Biomedicines

Article Title: REDD1 Affects Proliferation, Apoptosis, Migration, and Colony Formation via p-ERK and p-JNK Signaling in Lung Adenocarcinoma Cells Under Hypoxia

doi: 10.3390/biomedicines13122918

Figure Lengend Snippet: Knockdown of REDD1 inhibits A549 and H1299 cell proliferation under hypoxia. ( A , B ) A549 and H1299 cells were transfected with si-REDD1 under normoxic and hypoxic conditions. Then, the EdU assay was applied. ( C , D ) Quantitation of EdU + cells. ( E , F ) Cell viability was determined using the CCK-8 assay under the same conditions as in A. n = 4. Scale bar, 100 μm. * p < 0.05; ** p < 0.01.

Article Snippet: Antibodies that recognize REDD1 (10638-1-AP), matrix metallopeptidase 1 (MMP1) (10371-2-AP), matrix metallopeptidase 9 (MMP9) (60600-1-Ig), and B-cell lymphoma-2-associated X (Bax) (50599-2-Ig) were purchased from Proteintech (Wuhan, China).

Techniques: Knockdown, Transfection, EdU Assay, Quantitation Assay, CCK-8 Assay

Journal: Biomedicines

Article Title: REDD1 Affects Proliferation, Apoptosis, Migration, and Colony Formation via p-ERK and p-JNK Signaling in Lung Adenocarcinoma Cells Under Hypoxia

doi: 10.3390/biomedicines13122918

Figure Lengend Snippet: Downregulation of REDD1 promotes A549 and H1299 cell apoptosis under hypoxia. ( A , B ) Apoptosis of A549 and H1299 cells following the knockdown of REDD1 was detected by flow cytometric analysis of Annexin V-FITC/PI. ( C , D ) Quantitation of apoptotic cells. ( E , F ) Protein expression of Bax and Bcl2 was determined by Western blot. n = 4. * p < 0.05; ** p < 0.01.

Article Snippet: Antibodies that recognize REDD1 (10638-1-AP), matrix metallopeptidase 1 (MMP1) (10371-2-AP), matrix metallopeptidase 9 (MMP9) (60600-1-Ig), and B-cell lymphoma-2-associated X (Bax) (50599-2-Ig) were purchased from Proteintech (Wuhan, China).

Techniques: Knockdown, Quantitation Assay, Expressing, Western Blot

Journal: Biomedicines

Article Title: REDD1 Affects Proliferation, Apoptosis, Migration, and Colony Formation via p-ERK and p-JNK Signaling in Lung Adenocarcinoma Cells Under Hypoxia

doi: 10.3390/biomedicines13122918

Figure Lengend Snippet: Cell migration and colony formation were suppressed after REDD1 knockdown under hypoxia. ( A , B ) Images of crystal violet staining. ( C , D ) The number of migrating cells was analyzed. ( E – G ) Colony formation assay was performed in A549 and H1299 cells. n = 4. Scale bar, 100 μm. * p < 0.05; ** p < 0.01.

Article Snippet: Antibodies that recognize REDD1 (10638-1-AP), matrix metallopeptidase 1 (MMP1) (10371-2-AP), matrix metallopeptidase 9 (MMP9) (60600-1-Ig), and B-cell lymphoma-2-associated X (Bax) (50599-2-Ig) were purchased from Proteintech (Wuhan, China).

Techniques: Migration, Knockdown, Staining, Colony Assay

Journal: Biomedicines

Article Title: REDD1 Affects Proliferation, Apoptosis, Migration, and Colony Formation via p-ERK and p-JNK Signaling in Lung Adenocarcinoma Cells Under Hypoxia

doi: 10.3390/biomedicines13122918

Figure Lengend Snippet: Signaling pathway protein expression after REDD1 knockdown. ( A , B ) Western blot results for MAPK signaling proteins p-ERK, p-JNK, and p-p38 MAPK are shown in A549 and H1299 cells. ( C , D ) Identification of p-AKT and p-mTOR in A549 and H1299 cells. n = 4. * p < 0.05; ** p < 0.01.

Article Snippet: Antibodies that recognize REDD1 (10638-1-AP), matrix metallopeptidase 1 (MMP1) (10371-2-AP), matrix metallopeptidase 9 (MMP9) (60600-1-Ig), and B-cell lymphoma-2-associated X (Bax) (50599-2-Ig) were purchased from Proteintech (Wuhan, China).

Techniques: Expressing, Knockdown, Western Blot

Journal: Biomedicines

Article Title: REDD1 Affects Proliferation, Apoptosis, Migration, and Colony Formation via p-ERK and p-JNK Signaling in Lung Adenocarcinoma Cells Under Hypoxia

doi: 10.3390/biomedicines13122918

Figure Lengend Snippet: HN and ANI reverse the impact of the A549 cell line behavior. Cells were transfected with si-REDD1. ( A – D ) The EdU assay was used to measure cell proliferation. ( E , F ) Cell viability was determined by the CCK-8 test. ( G ) Annexin V-FITC/PI staining was used to evaluate cell apoptosis. ( H , I ) Images of cell migration are presented and quantified. n = 4. ANI, anisomycin, HN, honokiol, scale bar, 100 μm. * p < 0.05 and ** p < 0.01.

Article Snippet: Antibodies that recognize REDD1 (10638-1-AP), matrix metallopeptidase 1 (MMP1) (10371-2-AP), matrix metallopeptidase 9 (MMP9) (60600-1-Ig), and B-cell lymphoma-2-associated X (Bax) (50599-2-Ig) were purchased from Proteintech (Wuhan, China).

Techniques: Transfection, EdU Assay, CCK-8 Assay, Staining, Migration

Journal: Biomedicines

Article Title: REDD1 Affects Proliferation, Apoptosis, Migration, and Colony Formation via p-ERK and p-JNK Signaling in Lung Adenocarcinoma Cells Under Hypoxia

doi: 10.3390/biomedicines13122918

Figure Lengend Snippet: REDD1 regulates cell biological behavior in normoxic A549 and H1299 cells. Cells were transfected with REDD1 plasmid. Then, they were treated with U0126 (10 μM), SP600125 (10 μM), or SB203580 (10 μM) for 24 h under normoxia. ( A , B ) Cell proliferation was assessed by EdU assay. ( C , D ) The CCK-8 test was used to estimate cell viability. ( E , F ) Flow cytometry was used to detect apoptosis. ( G ) Cell migration and colony formation were presented using crystal violet staining. n = 4. * p < 0.05; ** p < 0.01.

Article Snippet: Antibodies that recognize REDD1 (10638-1-AP), matrix metallopeptidase 1 (MMP1) (10371-2-AP), matrix metallopeptidase 9 (MMP9) (60600-1-Ig), and B-cell lymphoma-2-associated X (Bax) (50599-2-Ig) were purchased from Proteintech (Wuhan, China).

Techniques: Transfection, Plasmid Preparation, EdU Assay, CCK-8 Assay, Flow Cytometry, Migration, Staining

Journal: iScience

Article Title: Single-cell transcriptomics of neuroinflammation and cerebrovascular endothelial cells in the aged rat hippocampus

doi: 10.1016/j.isci.2025.113332

Figure Lengend Snippet: Single-cell transcriptome analysis of the cerebral vascular endothelial cells (A–C) UMAP plot and bar plot showing the distribution of 6 subpopulations of cerebral vascular endothelial cells in the LPS and NC groups. (D) Violin plot shows the gene expression related to vascular origin (arterial, venous, and capillary), including arterial endothelial cell marker genes Fbln5, Bmx, Efnb2 , Vegfc . The venous endothelial cells highly expressed gene Nr2f and capillary endothelial cells highly expressed gene Rgcc and Slc16a1 . (E) Expression profiles of EC0 Marker genes including Mfge8, Lrg1, Lgals9, Cldn5, Ocln, Tjp1, Ddit4/Redd1, Mfsd2a are shown using the UMAP visualization approach. (F) Marker genes in the EC0 subpopulation are enriched with GO functional analysis. (G) Volcano plot depicts the DEGs at overall level of cerebral vascular endothelial cells between LPS and NC groups. DEGs (|log2(fold change)| > 1, p Value FDR <0.05, Difference = |pct.1- pct.2 | > 0.2) were colored (red for upregulated DEGs and blue for downregulated DEGs. (H and I) GO analysis shows the upregulated signaling pathway at overall level of cerebral vascular endothelial cells and EC0 subpopulation respectively.

Article Snippet: REDD1 specific antibody , Proteintech , Cat# 10638-1-AP; RRID: AB_2245711.

Techniques: Gene Expression, Marker, Expressing, Functional Assay

Journal: iScience

Article Title: Single-cell transcriptomics of neuroinflammation and cerebrovascular endothelial cells in the aged rat hippocampus

doi: 10.1016/j.isci.2025.113332

Figure Lengend Snippet: Cerebral vascular endothelial cells REDD1 expression induced by LPS exposure (A) Representative immunofluorescence double staining images of REDD1 (red) and CD31 (green) in the aging rat hippocampus on day 3 after a single intraperitoneal injection of LPS. Scale bar = 25 μm. Quantitative analysis of REDD1 fluorescence in endothelial cells (CD31 + ). (B) Representative confocal microscopic images of NC and LPS rats hippocampus stained for IgG. Nuclei are labeled with DAPI. Scale bars: 75 μm or 25 μm. Quantitative analysis of IgG extravascular deposits in two groups at hippocampus DG region. (C) Representative Western blot bands and quantitative analysis of REDD1 protein in hCMEC/D3 cells stimulated by LPS at an intervening concentration of 100 ng/mL for 24 h ( p < 0.0001). (D) Changes in REDD1 mRNA expression were measured by qPCR in hCMEC/D3 cells stimulated by LPS at an intervening concentration of 100 ng/mL for 24 h. Data are expressed as mean ± SEM, independent samples t-test, n ≥ 3, ∗∗ p < 0.01, ∗∗∗∗ p < 0.0001.

Article Snippet: REDD1 specific antibody , Proteintech , Cat# 10638-1-AP; RRID: AB_2245711.

Techniques: Expressing, Immunofluorescence, Double Staining, Injection, Fluorescence, Staining, Labeling, Western Blot, Concentration Assay

Journal: iScience

Article Title: Single-cell transcriptomics of neuroinflammation and cerebrovascular endothelial cells in the aged rat hippocampus

doi: 10.1016/j.isci.2025.113332

Figure Lengend Snippet: REDD1 gene knock-down decreased oxidative stress related cell damage and apoptosis (A and B) ROS production assessed by DHE (red) staining of hCMEC/D3 cells after interventions. Nuclei are labeled with DAPI. Relative intracellular DHE fluorescence among four groups: PBS+shNC, PBS+shREDD1, LPS+shNC, and LPS+shREDD1 group. Scale bar = 50 μm. (C) Relative fluorescence density of intracellular TMRM (red). (D) Relative ATP production in hCMEC/D3 cells after LPS stimulation and gene transfection. (E and F) Representative images of the fluorescence assay (left panel) and quantification (right panel) of double-staining of TUNEL (green) and DAPI (blue) on hCMEC/D3 cells among four groups. Scale bars = 50 μm. (G) The hCMEC/D3 cells supernatants were collected for LDH release assays. Data are expressed as mean ± SEM, one-way ANOVA with Tukey’s test, n ≥ 3, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗∗ p < 0.0001. Data represent three experiments for in vitro assays.

Article Snippet: REDD1 specific antibody , Proteintech , Cat# 10638-1-AP; RRID: AB_2245711.

Techniques: Knockdown, Staining, Labeling, Fluorescence, Transfection, Double Staining, TUNEL Assay, In Vitro