Journal: International journal of hyperthermia : the official journal of European Society for Hyperthermic Oncology, North American Hyperthermia Group

Article Title: Hyperthermia promotes exosome secretion by regulating Rab7b while increasing drug sensitivity in adriamycin-resistant breast cancer.

doi: 10.1080/02656736.2022.2029585

Figure Lengend Snippet: Figure 4. The validation experiment of hub genes. (A) The differences in gene expression of 10 hub genes were validated with RT-qPCR. (B) Western blot analysis of FOS, CREB5, MAPK8 and NFKB1 protein level. (C) The efficiency of siRNA to knockdown the expression of FOS and CREB5. (D) Cell proliferation in the si-FOS and si-CREB5 group was faster compared with that in the control group, using the CCK-8 assay. (E). Half-inhibition rate of adriamycin in MCF-7/ADR cells treated with si-FOS and si-CREB5. (F) Different expression of FOS and CERB5 between invasive breast cancer and normal breast tissue by TCGA database. Error bars represented the mean ± SD of at least three independent experiments, p < .05, p < .01, p < .001.

Article Snippet: The membrane was then probed with primary antibodies at 4 C overnight and with species-specific secondary antibodies at room temperature for 1 h. The primary antibodies, including FOS antibody (1:1000, Proteintech, 66590-1-Ig), NF-jB antibody (1:1000, Proteintech, 14220-1-AP), MAPK8 antibody (1:1000, Proteintech, 66210-1-Ig), CREB5 antibody (1:1000, Proteintech, 14196-1-AP), GAPDH antibody (1:5000, Proteintech, 60004-1-Ig), CD9 antibody (1:1000, Abcam, ab223052), CD63 antibody (1:1000, Abcam, ab68418), TSG101 Figure 1.

Techniques: Biomarker Discovery, Gene Expression, Quantitative RT-PCR, Western Blot, Knockdown, Expressing, Control, CCK-8 Assay, Inhibition

Journal: International journal of hyperthermia : the official journal of European Society for Hyperthermic Oncology, North American Hyperthermia Group

Article Title: Hyperthermia promotes exosome secretion by regulating Rab7b while increasing drug sensitivity in adriamycin-resistant breast cancer.

doi: 10.1080/02656736.2022.2029585

Figure Lengend Snippet: Figure 5. Hyperthermia promotes the secretion of exosome in MCF-7/ADR cells. (A) The expression level of FOS and CREB5 in MCF-7/ADR exosomes after hyper- thermia. (B) Half-inhibition rate of adriamycin in MCF-7/ADR cells incubated with the exosomes which produced by MCF-7/ADR cells after hyperthermia. (C) The expression level of FOS and CREB5 in MCF-7/ADR after incubated with hyperthermia ADR/exo. (D) Particles concentration of exosomes derived from breast cancer cells analyzed by NTA. (E) Total protein amounts of exosomes derived from breast cancer cells analyzed by bicinchoninic acid (BCA) protein kit. (F) Transmission electron micrographs (TEM) of cells before or 6 h after hyperthermia. Intense extracellular vesicle shedding occurred 6 h after hyperthermia. Scale bar ¼1 lm. (G) Confocal microscope analysis of exosome uptake by MCF-7/ADR cells with or without hyperthermia treatment. Scale bar ¼20 lm. Error bars represent the mean ± SD of at least three independent experiments, p < .05, p < .01, p < .001.

Article Snippet: The membrane was then probed with primary antibodies at 4 C overnight and with species-specific secondary antibodies at room temperature for 1 h. The primary antibodies, including FOS antibody (1:1000, Proteintech, 66590-1-Ig), NF-jB antibody (1:1000, Proteintech, 14220-1-AP), MAPK8 antibody (1:1000, Proteintech, 66210-1-Ig), CREB5 antibody (1:1000, Proteintech, 14196-1-AP), GAPDH antibody (1:5000, Proteintech, 60004-1-Ig), CD9 antibody (1:1000, Abcam, ab223052), CD63 antibody (1:1000, Abcam, ab68418), TSG101 Figure 1.

Techniques: Expressing, Inhibition, Incubation, Produced, Concentration Assay, Derivative Assay, Transmission Assay, Microscopy

Journal: Life sciences in space research

Article Title: Comparison of signaling profiles in the low dose range following low and high LET radiation.

doi: 10.1016/j.lssr.2020.02.002

Figure Lengend Snippet: Fig. 1. Average total fold intensity of phospho-protein signal as compared to median control value. Colored bars (dark blue = 0.5 Gy, light blue = 0.1 Gy and green = 0.05 Gy) indicate values significantly different from controls (yellow). Significance bars are shown for all significant differences between doses and 0 (p ≤0.05). Average fold intensity levels are shown at 2 h post radiation for γH2AX (A), pATF2 (B) and pSMC1 (C). Persistent effects are shown at 24 h for γH2AX (D), pATF2 (E) and pSMC1 (F). (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: For staining, 0.5 × 106 fixed cells were washed in phosphate buffered saline (PBS), resuspended in blocking buffer (2% FBS/PBS) and incubated for 1 h with primary antibody on ice, with resuspension of the pellet every 15 min. Primary antibodies used in flow cytometry include mouse monoclonal γH2AXSer139 (1:800 dilution) and pSMC1Ser957 (1:800 dilution) from Millipore (Temecula, CA) and rabbit polyclonal pATF2 Ser490/498 (1:1000 dilution) from Rockland, Inc. (Gilbertsville, PA).

Techniques: Control

Journal: Life sciences in space research

Article Title: Comparison of signaling profiles in the low dose range following low and high LET radiation.

doi: 10.1016/j.lssr.2020.02.002

Figure Lengend Snippet: Fig. 3. Average total fold pATF2 intensity over median control level versus fluence for various radiation qualities. Average fold intensity levels are shown at 2 h post radiation for Si ions (A),Fe ions (C) and Ti ions (E). Persistent effects are shown at 24 h for Si ions (B), Fe ions (D) and Ti ions (F).

Article Snippet: For staining, 0.5 × 106 fixed cells were washed in phosphate buffered saline (PBS), resuspended in blocking buffer (2% FBS/PBS) and incubated for 1 h with primary antibody on ice, with resuspension of the pellet every 15 min. Primary antibodies used in flow cytometry include mouse monoclonal γH2AXSer139 (1:800 dilution) and pSMC1Ser957 (1:800 dilution) from Millipore (Temecula, CA) and rabbit polyclonal pATF2 Ser490/498 (1:1000 dilution) from Rockland, Inc. (Gilbertsville, PA).

Techniques: Control

Journal: Life sciences in space research

Article Title: Comparison of signaling profiles in the low dose range following low and high LET radiation.

doi: 10.1016/j.lssr.2020.02.002

Figure Lengend Snippet: Fig. 6. Average total fold intensity of pATF2 signal intensity divided by fluence and graphed versus LET. Average fold intensity levels are shown at 2 h post radiation for 0.05 Gy (A), 0.1 Gy (B) and 0.5 Gy (C). Persistent effects are shown at 24 h for 0.05 Gy (D), 0.1 Gy (E) and 0.5 Gy (F).

Article Snippet: For staining, 0.5 × 106 fixed cells were washed in phosphate buffered saline (PBS), resuspended in blocking buffer (2% FBS/PBS) and incubated for 1 h with primary antibody on ice, with resuspension of the pellet every 15 min. Primary antibodies used in flow cytometry include mouse monoclonal γH2AXSer139 (1:800 dilution) and pSMC1Ser957 (1:800 dilution) from Millipore (Temecula, CA) and rabbit polyclonal pATF2 Ser490/498 (1:1000 dilution) from Rockland, Inc. (Gilbertsville, PA).

Techniques:

Journal: Cell reports

Article Title: Functional analyses of human LUC7-like proteins involved in splicing regulation and myeloid neoplasms

doi: 10.1016/j.celrep.2021.108989

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: The knockdowns were confirmed by testing the expression levels of the LUC7-like family by western blot using anti-LUC7L (Protein Tech, 17085-1-AP), anti-LUC7L2 (Bethyl, A304-504A-M), and anti-LUC7L3 (Protein Tech, 14504-1-AP).

Techniques: Recombinant, Isolation, Reverse Transcription, Knockdown, Mass Spectrometry, shRNA, Plasmid Preparation, Software

Journal: Scientific Reports

Article Title: Effects of ATF2/TSC1 on epilepsy by modulating the microphages polarization of microglia

doi: 10.1038/s41598-025-04914-4

Figure Lengend Snippet: ATF2 is overexpressed in KA-induced HHMC3 cells. ( A ) ATF2 mRNA expression in KA-induced HMC3 cell. ( B - C ) ATF2 protein expression in KA-induced HMC3 cell. ( D ). The fluorescence intensity of ATF2 and IBA1. Analysis of the relative fluorescence intensity of ATF2 ( E ) and IBA1 ( F ). * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Cells were blocked with 3% bovine serum albumin for 30 min and incubated with IBA1 (Proteintech, China; 1: 500) and ATF2 (Proteintech, China; 1: 100) primary antibody at 4 °C overnight.

Techniques: Expressing, Fluorescence

Journal: Scientific Reports

Article Title: Effects of ATF2/TSC1 on epilepsy by modulating the microphages polarization of microglia

doi: 10.1038/s41598-025-04914-4

Figure Lengend Snippet: ATF2 inhibition represses the M1 polarization of HMC3 cells. ( A - B ) ATF2 protein level in HMC3 cells transfected with sh-ATF2. ( C - D ) ATF2 protein expression in KA-induced HMC3 cells transfected with sh-ATF2. Flow cytometry was performed to access cell ratio of M1 microphage CD80 ( E - F ), IL-1β ( G - H ) and M2 microphage marker CD206 ( I - J ), CD163 ( K - L ). * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Cells were blocked with 3% bovine serum albumin for 30 min and incubated with IBA1 (Proteintech, China; 1: 500) and ATF2 (Proteintech, China; 1: 100) primary antibody at 4 °C overnight.

Techniques: Inhibition, Transfection, Expressing, Flow Cytometry, Marker

Journal: Scientific Reports

Article Title: Effects of ATF2/TSC1 on epilepsy by modulating the microphages polarization of microglia

doi: 10.1038/s41598-025-04914-4

Figure Lengend Snippet: ATF2 inhibition represses the M1 polarization of HMC3 cells. The mRNA expression of TNF-α ( I ), IL-6 ( B ), TGF-β ( C ), and IL-10 ( D ) in HMC3 cells transfected with sh-ATF2. ( E ) The gel images of INOS and ARG1. ( F ) Relative expression of INOS protein. ( G ) Relative expression of ARG1 protein. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Cells were blocked with 3% bovine serum albumin for 30 min and incubated with IBA1 (Proteintech, China; 1: 500) and ATF2 (Proteintech, China; 1: 100) primary antibody at 4 °C overnight.

Techniques: Inhibition, Expressing, Transfection

Journal: Scientific Reports

Article Title: Effects of ATF2/TSC1 on epilepsy by modulating the microphages polarization of microglia

doi: 10.1038/s41598-025-04914-4

Figure Lengend Snippet: ATF2 regulates TSC1 transcription. ( A ) The binding sites of ATF2 obtained from JASPAR database. ( B ) The promoter regions (P1: promoter region 1, P2: promoter region 2, P3: promoter region 3) of TSC1. ( C ) The relative enrichment of ATF2 in TSC1 promoter regions. Dual luciferase reporter ( D ) and western blot ( E ) assay revealed that ATF2 inhibited the transcription of TSC1. ( F ) The luciferase activity of wild-type (WT) and mutant (MUT) TSC1 in HMC3 cells. ( G ) Relative expression of TSC1 in KA-induced HMC3 cells. ( H - I ) Transfection with sh-ATF2 enhanced TSC1 protein. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Cells were blocked with 3% bovine serum albumin for 30 min and incubated with IBA1 (Proteintech, China; 1: 500) and ATF2 (Proteintech, China; 1: 100) primary antibody at 4 °C overnight.

Techniques: Binding Assay, Luciferase, Western Blot, Activity Assay, Mutagenesis, Expressing, Transfection

Journal: Scientific Reports

Article Title: Effects of ATF2/TSC1 on epilepsy by modulating the microphages polarization of microglia

doi: 10.1038/s41598-025-04914-4

Figure Lengend Snippet: ATF2 regulates microglia polarization in KA-induced mouse by medicating TSC1 transcription. ( A ) Racine scale scores were used to evaluate the seizure severity of mice. ( B ) HE staining of mouse hippocampal tissues. ( C - D ) Nissl staining of mouse hippocampal tissues. ( E ) The gel images of ATF2 and TSC1. The relative expression of ATF2 ( F ) and TSC1 ( G ). ( H - I ) The fluorescence staining of IBA1 and CD80. ( J - K ) The fluorescence staining of IBA1 and CD206. The relative expression of TNF-α ( L ), IL-6 ( M ), TGF-β ( N ), and IL-10 ( O ). * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Cells were blocked with 3% bovine serum albumin for 30 min and incubated with IBA1 (Proteintech, China; 1: 500) and ATF2 (Proteintech, China; 1: 100) primary antibody at 4 °C overnight.

Techniques: Staining, Expressing, Fluorescence

Journal: iScience

Article Title: CREB3L4 promotes hepatocellular carcinoma progression and decreases sorafenib chemosensitivity by promoting RHEB-mTORC1 signaling pathway

doi: 10.1016/j.isci.2024.108843

Figure Lengend Snippet:

Article Snippet: CREB3L4 , Proteintech , Cat No. 13630-1-AP; RRID: AB_2276550.

Techniques: Virus, Recombinant, CCK-8 Assay, Chromatin Immunoprecipitation, Reporter Assay, Expressing, Sequencing, Plasmid Preparation, Software