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: Development (Cambridge, England)

Article Title: The Wnt/JNK signaling target gene alcam is required for embryonic kidney development.

doi: 10.1242/dev.107938

Figure Lengend Snippet: Fig. 7. The Fzd3-responsive element (Fzd3RE) is regulated through Pax2 and ATF2. (A) ATF2 and Pax2 bind to the −1752 to −2013 region, as shown by chromatin immunoprecipitation using a primer pair specific for the Fzd3RE (Fzd3RE primer). A primer pair matching part of the coding region of alcam serves as negative control (control primer). +ab, immunoprecipitation using either ATF2 or Pax2 antibody, as indicated; −ab, control reaction in the absence of antibody. Binding of ATF2 or Pax2 to the Fzd3RE is dependent on Fzd3 signaling, as shown by considerably weaker signals in the Fzd3 MO reactions. (B) The −2.7 kb-luc reporter responds to co-transfection of pax2. This regulation depends on the −1752 to −2013 region. (C) Downregulation of alcam expression upon loss of Fzd3 (arrow) is rescued by co-injecting pax2 RNA. Lateral views with anterior towards the left (injected side) or towards the right (uninjected side) are shown. A quantitative representation of the experiment is shown. n, number of independent experiments; N, number of analyzed embryos in total; luc, luciferase reporter gene; RLU, relative light units. *P≤0.05. (D) Different β-Catenin- independent Wnt signaling branches are required for tubular morphogenesis. Left (blue): the Wnt/ROCK signaling pathway results in cytoskeletal rearrangement. Right (red): Wnt/JNK signaling directly activates alcam gene transcription involving ATF2 [as a heterodimer with other yet unknown bZIP (basic zipper) transcription factors] and Pax2.

Article Snippet: Immunoprecipitation was performed with ATF2 (Cell Signaling) or Pax2 antibodies (Abcam).

Techniques: Chromatin Immunoprecipitation, Negative Control, Control, Immunoprecipitation, Binding Assay, Cotransfection, Expressing, Injection, Luciferase

Journal: iScience

Article Title: THBS1 upregulation by KLF7 in hypopharyngeal squamous cell carcinoma contributes to lung metastasis through the p38 MAPK signaling

doi: 10.1016/j.isci.2026.114816

Figure Lengend Snippet: The p38 MAPK activator LX-3 rescues the malignant behavior of HPSCC cells repressed by the silencing of THBS1 (A) Western blot analysis of the extent of p38, ATF2, and MK2 phosphorylation, as well as total p38 expression in KD-THBS1-treated HPSCC cells. (B) Western blot analysis of the extent of p38, ATF2, and MK2 phosphorylation, as well as total p38 expression in HPSCC cells treated with 2 μM LX-3 after knockdown of THBS1. (C–E) HPSCC cell proliferative capacity was examined using EdU staining. Transwell assay detects changes in the migratory (D) and invasive (E) abilities of HPSCC cells. (F) Expression of E-cadherin and N-cadherin in HPSCC cells was examined using dual-labeling immunofluorescence. The results are expressed as the mean ± SD (A–F). These values are derived from three independent experiments. Statistical analysis was performed using an unpaired t test.

Article Snippet: The primary antibodies used were as follows: anti-THBS1 (1:250, A2125, BioAb), anti-E-cadherin (1:1000, 3195, Cell Signaling Technologies), anti-N-cadherin (1:100, PA5-19486m Invitrogen), anti-Phospho-p38 MAPK (Thr180/Tyr182) (1:1000, 4511, Cell Signaling Technologies), anti-total p38 (1:1000, 9212, Cell Signaling Technologies), anti- p -ATF2 (Thr71) (1:1000, 44-295G, Invitrogen), anti-p-MK2 (Thr334) (1:1000, 3007, Cell Signaling Technologies), anti-KLF7 (1:1000, CSB- PA126129 , Cusabio), anti-Slug (1:1000, 12129-1-AP, ProteinTech Group), anti-Vimentin (1:20000, 10366-1-AP, ProteinTech Group), anti-HDAC6 (1:1000, 12834-1-AP, ProteinTech Group), and anti-GAPDH (1:20000, AC001, BioAb).

Techniques: Western Blot, Phospho-proteomics, Expressing, Knockdown, Staining, Transwell Assay, Labeling, Immunofluorescence, Derivative Assay

Journal: iScience

Article Title: THBS1 upregulation by KLF7 in hypopharyngeal squamous cell carcinoma contributes to lung metastasis through the p38 MAPK signaling

doi: 10.1016/j.isci.2026.114816

Figure Lengend Snippet: Reactivation of THBS1 rescues the HPSCC cell malignant biological behavior curtailed by knockdown of KLF7 (A) Western blot analysis of protein expression of KLF7 and THBS1 in HPSCC cells treated with knockdown of KLF7 combined with the overexpression of THBS1. (B–D) HPSCC cell proliferative capacity was examined using EdU staining. Transwell assay detects changes in the migratory (C) and invasive (D) abilities of HPSCC cells. (E) Expression of E-cadherin and N-cadherin in HPSCC cells was examined using dual-labeling immunofluorescence. (F) Western blot analysis of the extent of p38, ATF2, and MK2 phosphorylation, as well as total p38 expression in HPSCC cells. The results are expressed as the mean ± SD (A–F). These values are derived from three independent experiments. Statistical analysis was performed using a one-way ANOVA followed by Tukey’s multiple comparison test.

Article Snippet: The primary antibodies used were as follows: anti-THBS1 (1:250, A2125, BioAb), anti-E-cadherin (1:1000, 3195, Cell Signaling Technologies), anti-N-cadherin (1:100, PA5-19486m Invitrogen), anti-Phospho-p38 MAPK (Thr180/Tyr182) (1:1000, 4511, Cell Signaling Technologies), anti-total p38 (1:1000, 9212, Cell Signaling Technologies), anti- p -ATF2 (Thr71) (1:1000, 44-295G, Invitrogen), anti-p-MK2 (Thr334) (1:1000, 3007, Cell Signaling Technologies), anti-KLF7 (1:1000, CSB- PA126129 , Cusabio), anti-Slug (1:1000, 12129-1-AP, ProteinTech Group), anti-Vimentin (1:20000, 10366-1-AP, ProteinTech Group), anti-HDAC6 (1:1000, 12834-1-AP, ProteinTech Group), and anti-GAPDH (1:20000, AC001, BioAb).

Techniques: Knockdown, Western Blot, Expressing, Over Expression, Staining, Transwell Assay, Labeling, Immunofluorescence, Phospho-proteomics, Derivative Assay, Comparison

Journal: iScience

Article Title: THBS1 upregulation by KLF7 in hypopharyngeal squamous cell carcinoma contributes to lung metastasis through the p38 MAPK signaling

doi: 10.1016/j.isci.2026.114816

Figure Lengend Snippet: Activation of the THBS1/p38 MAPK axis by KLF7 promotes the lung metastasis of HPSCC cells (A) Lung metastasis in nude mice injected with FaDu cells in the tail vein was assessed by HE staining. (B–D) Quantitative results of lung metastatic nodules. IHC scores (C) and protein expression (D) of Slug and Vimentin within lung metastases in nude mice. (E) RT-qPCR detection of mRNA expression levels of KLF7 and THBS1 in lung metastases. (F) IHC scores of KLF7 and THBS1 within lung metastases in nude mice. (G) Western blot analysis of the extent of p38, ATF2, and MK2 phosphorylation, as well as total p38 expression in lung metastases. The results are expressed as the mean ± SD (A–G). These values are derived from five mice. Statistical analysis was performed using a one-way ANOVA followed by Tukey’s multiple comparison test.

Article Snippet: The primary antibodies used were as follows: anti-THBS1 (1:250, A2125, BioAb), anti-E-cadherin (1:1000, 3195, Cell Signaling Technologies), anti-N-cadherin (1:100, PA5-19486m Invitrogen), anti-Phospho-p38 MAPK (Thr180/Tyr182) (1:1000, 4511, Cell Signaling Technologies), anti-total p38 (1:1000, 9212, Cell Signaling Technologies), anti- p -ATF2 (Thr71) (1:1000, 44-295G, Invitrogen), anti-p-MK2 (Thr334) (1:1000, 3007, Cell Signaling Technologies), anti-KLF7 (1:1000, CSB- PA126129 , Cusabio), anti-Slug (1:1000, 12129-1-AP, ProteinTech Group), anti-Vimentin (1:20000, 10366-1-AP, ProteinTech Group), anti-HDAC6 (1:1000, 12834-1-AP, ProteinTech Group), and anti-GAPDH (1:20000, AC001, BioAb).

Techniques: Activation Assay, Injection, Staining, Expressing, Quantitative RT-PCR, Western Blot, Phospho-proteomics, Derivative Assay, Comparison