Journal: Frontiers in Molecular Neuroscience

Article Title: Evaluation of the amyloid beta-GFP fusion protein as a model of amyloid beta peptides-mediated aggregation: a study of DNAJB6 chaperone

doi: 10.3389/fnmol.2015.00040

Figure Lengend Snippet: Canonical members of HSPB family prevent the aggregation of Aβ-GFP. Cells were transfected with HSPB1, HSPB5, HSPB7 or FRTTO at 1:3 ratio for 48 h. Quantification of the pellet/soluble ratio of Aβ-GFP relative to FRTTO was depicted in the chart above the blot. Values represent mean ± SE of two independent experiments. ** P < 0.01, ns = non significant.

Article Snippet: The membranes were blocked with 5% dry milk in PBS with 0.1% Tween 20 (PBST) for 1 h at room temperature and incubated overnight at 4°C with the following primary antibodies: 6E10 (1:1000 in TBST, Covance), anti V5 (1:5000 in PBST, Invitrogen), anti β-actin (1:1000 in PBST, Abcam), anti HSPB1 (1:1000 in PBST, Stress Marq Biosciences), anti HSPB5 (1:1000 in PBST, Stress Marq Biosciences) and anti HSPB7 (1:1000, Abnova).

Techniques: Transfection

Journal: Journal for immunotherapy of cancer

Article Title: Vinblastine resets tumor-associated macrophages toward M1 phenotype and promotes antitumor immune response.

doi: 10.1136/jitc-2023-007253

Figure Lengend Snippet: Figure 4 VBL targets NF-κB-Cyba to reprogram macrophages. (A) Gene ontology analysis of RNA-sequencing data collected from BMDMs with or without VBL treatment for 24 hours. (B) A heatmap of genes related to the NFkB pathway from RNA- sequencing data collected from macrophages (Mϕ in vivo), sorted and analyzed from Vehicle/VBL treated tumors. The heat map was created based on the normalized FPKM of genes in the NF-κB pathway. (C) Venn diagrams of upregulated genes enriched in ROS production pathway. (D) Gene expression of Mmp8, Cyba, Cd36, Dcxr, and Alox5 in macrophages sorted from MC38 tumors. n=4. (E) Representative western blot against p-NF-κB, NF-κB, p22phox, and β-actin in macrophages sorted from indicated tumor samples. (F) Fold change of p-NF-κB, NF-κB, and p22phox protein level in panel (E). n=3. (G) Representative western blot against p22phox and β-actin in BMDMs. (H) Fold change of p22phox protein levels in panel (G). n=3. (I) Flow cytometry analysis for percentages of M1 (F4/80+CD11b+CD86+) and M2 (F4/80+CD11b+CD206+) BMDMs after the corresponding treatment for 24 hours. n=6. (J) Representative flow cytometry results of T cell proliferation after co-culture with BMDMs for 72 hours. (K) Statistical results of T cell proliferation in panel (J). n=6. Data are presented as mean±SD. P values were determined by a two-tailed Student’s t-test. *p<0.05, **p<0.01, ***p<0.001. BMDMs, bone marrow-derived macrophages; ROS, reactive oxygen species; VBL, vinblastine.

Article Snippet: The following commercial antibodies were used: antiphospho- NF-κB (Cell Signaling Technology, 3033S), antiNF-κB (Cell Signaling Technology, 8242S), anti- p22phox (Cell Signaling Technology, 27 297S), anti-β-actin (Sigma A1978), anti- mouse (Abcam ab6728), anti- Rabbit (Abcam ab6721).

Techniques: RNA Sequencing, In Vivo, Gene Expression, Western Blot, Flow Cytometry, Co-Culture Assay, Two Tailed Test, Derivative Assay

Journal: Journal for immunotherapy of cancer

Article Title: Vinblastine resets tumor-associated macrophages toward M1 phenotype and promotes antitumor immune response.

doi: 10.1136/jitc-2023-007253

Figure Lengend Snippet: Figure 8 Working model for VBL-mediated antitumor immune response. VBL phosphorylates and activates the transcription factor NF-κB in TAMs, upregulating cyba which encodes p22phox protein expression. Cytosolic ROS and MitoROS levels were increased by p22phox -catalyzed synthesis. ROS further facilitated the entry of transcription factor TFEB into the nucleus, accelerating lysosome biogenesis and activation, and thus endowing TAMs with a higher phagocytic capacity. ROS also repolarized TAMs into the M1-like phenotype and abolished their suppressive activity on CD8+ T cells. This graph model was created with biorender.com. ROS, reactive oxygen species; TAMs, tumor-associated macrophage; VBL, vinblastine.

Article Snippet: The following commercial antibodies were used: antiphospho- NF-κB (Cell Signaling Technology, 3033S), antiNF-κB (Cell Signaling Technology, 8242S), anti- p22phox (Cell Signaling Technology, 27 297S), anti-β-actin (Sigma A1978), anti- mouse (Abcam ab6728), anti- Rabbit (Abcam ab6721).

Techniques: Expressing, Activation Assay, Activity Assay

Journal: Immunity

Article Title: Quantitative control of Ets1 dosage by a multienhancer hub promotes Th1 cell differentiation and protects from allergic inflammation

doi: 10.1016/j.immuni.2023.05.004

Figure Lengend Snippet: Key resources table

Article Snippet: Anti-Acetyl-Histone H3 (Lys27) (D5E4) , Cell Signaling Technology , Cat# 8173; RRID: AB_10949503.

Techniques: Recombinant, Staining, Flow Cytometry, Protease Inhibitor, Microscopy, Electron Microscopy, Purification, Cell Isolation, Gene Expression, Plasmid Preparation, Software