Journal: Cancer Immunology Research

Article Title: Tumor Cell–Intrinsic Decr2 Regulates Ferroptosis and Immunotherapy Efficacy

doi: 10.1158/2326-6066.CIR-24-0519

Figure Lengend Snippet: Decr2 expression correlates with patient outcomes, and Slc7a11 is a candidate therapeutic target. A, Fold changes of Decr2 in matched pretherapy and on-therapy samples from patients with melanoma. B, Relationship between overall survival and mRNA expression level of Decr2 in human patients treated with ipilimumab. C, Decr2 deletion (deep and shallow) and Decr2 copy-number gain association with overall survival in response to ipilimumab. D, IF staining of human melanoma tissue sections. DECR2 (green), SOX10 (red), and DAPI (blue). Scale bar, 50 μm. E, Percentage of Decr2 + Sox10 + cells and relative intensity of DECR2 expression in tissues of patients with melanoma: pretreatment or on-treatment with anti–PD-1. F, KD of Slc7a11 in Decr2 -deficient tumor cells restores anti–PD-L1 efficacy. Control shRNA–infected tumors (in black), Decr2 single KD tumors (in red, labeled as Decr2 KD), and Decr2 Slc7a11 double KD (in blue, labeled as Decr2 Slc7a11 DKD). n ≥ 9 mice per group. G, The end timepoint of tumor area with or without anti–PD-L1 treatment. Control shRNA–infected tumors (in black), Decr2 single KD tumors (in red, labeled as Decr2 KD), Decr2 Slc7a11 double KD (in blue, labeled as Decr2 Slc7a11 DKD). P values are determined using the Mann–Whitney test ( A ), the two-tailed t test ( C, E, G ), and two-way ANOVA ( F ). *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001; ns, not significant. CR, complete responder; PD, progressive disease; PR, partial responder; SD, stable disease.

Article Snippet: An IF protocol was developed using antibodies specific to DECR2 (Abcam, ab153849, 1/50 dilution) and SOX10 (R&D Systems, MAB2864, 1/200 dilution) on formalin-fixed, paraffin-embedded tissue sections.

Techniques: Expressing, Staining, Control, shRNA, Infection, Labeling, MANN-WHITNEY, Two Tailed Test

Journal: Pharmaceutical Biology

Article Title: Baicalein attenuates bleomycin-induced lung fibroblast senescence and lung fibrosis through restoration of Sirt3 expression

doi: 10.1080/13880209.2022.2160767

Figure Lengend Snippet: Baicalein mitigates BLM-induced senescence in lung tissues and isolated lung fibroblasts. (A, B) Protein levels of p53, p21 and p16 and the mRNA levels of SASP factors, including MCP-1, PAI-1, TNF-α, MMP-10 and MMP-12 in lung tissues were measured in control, BLM, baicalein and BLM + baicalein groups. (C, D) Protein levels of p53, p21 and p16 and the mRNA levels of MCP-1, PAI-1, TNF-α, MMP-10 and MMP-12 in isolated lung fibroblasts were measured using western blotting and RT-qPCR, respectively. Representative protein bands were presented on the top of the histograms (A, C). Data are presented as the mean ± SEM ( n = 7). ** p < 0.01 vs. control. ## p < 0.01 vs. BLM. SASP: senescence-associated secretory phenotype; MCP-1: monocyte chemotactic protein-1; PAI-1: plasminogen activator inhibitor-1; TNF-α: tumour necrosis factor-α; MMP: matrix metalloproteinase; RT-qPCR: reverse transcription-quantitative PCR; BLM: bleomycin.

Article Snippet: Following blocking with non-fat dry milk dissolved in TBS for 2 h at room temperature, the membranes were incubated with BSA-diluted primary antibodies against Sirt3 (1:1000; cat. no. sc-365175; Santa Cruz Biotechnology, Inc., Dallas, TX), p16 (1:1000, cat. no. sc-56330; Santa Cruz Biotechnology, Inc., Dallas, TX), Smad4 (1:1000; cat. no. sc-7966; Santa Cruz Biotechnology, Inc., Dallas, TX), p-Smad2 (p-Smad2; 1:1000; Cell Signaling Technology, Inc., Danvers, MA), p-Smad3 (1:1000; Cell Signaling Technology, Inc., Danvers, MA), p53 (1:1000; cat. no. 10422-1-AP; ProteinTech Group, Inc., Rosemont, IL), p21 (1:1000; cat. no. 27296-1-AP; ProteinTech Group, Inc., Rosemont, IL), αSMA (1:1000, cat. no. sc-8432; Santa Cruz Biotechnology, Inc., Dallas, TX), fibronectin (1:1000, cat. no. sc-8422; Santa Cruz Biotechnology, Inc., Dallas, TX), Smad2 (1:1000; cat. no. sc-393312 Santa Cruz Biotechnology, Inc., Dallas, TX), Smad3 (1:1000; cat. no. sc-101154; Santa Cruz Biotechnology, Inc., Dallas, TX) or β-actin (1:1000; cat. no. sc-81178; Santa Cruz Biotechnology, Inc., Dallas, TX) at 4 °C overnight.

Techniques: Isolation, Control, Western Blot, Quantitative RT-PCR, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Pharmaceutical Biology

Article Title: Baicalein attenuates bleomycin-induced lung fibroblast senescence and lung fibrosis through restoration of Sirt3 expression

doi: 10.1080/13880209.2022.2160767

Figure Lengend Snippet: Sirt3 silencing abolishes the protective effect of baicalein against BLM-induced senescence in lung tissue and isolated lung fibroblasts. (A, B, D and E) The protein expression levels of Sirt3, p53, p21 and p16 in pulmonary tissues were measured using western blotting. (C, F) The mRNA levels of MCP-1, PAI-1, TNF-α, MMP-10 and MMP-12 were measured using RT-qPCR. Data are presented as the mean ± SEM ( n = 7). ** p < 0.01 vs. control. ## p < 0.01 vs. BLM. $$ p < 0.01 vs. BLM + baicalein. Sirt3: sirtuin 3; BLM: bleomycin; MCP-1: monocyte chemotactic protein-1; PAI-1: plasminogen activator inhibitor-1; TNF-α: tumour necrosis factor-α; MMP: matrix metalloproteinase; RT-qPCR: reverse transcription-quantitative PCR.

Article Snippet: Following blocking with non-fat dry milk dissolved in TBS for 2 h at room temperature, the membranes were incubated with BSA-diluted primary antibodies against Sirt3 (1:1000; cat. no. sc-365175; Santa Cruz Biotechnology, Inc., Dallas, TX), p16 (1:1000, cat. no. sc-56330; Santa Cruz Biotechnology, Inc., Dallas, TX), Smad4 (1:1000; cat. no. sc-7966; Santa Cruz Biotechnology, Inc., Dallas, TX), p-Smad2 (p-Smad2; 1:1000; Cell Signaling Technology, Inc., Danvers, MA), p-Smad3 (1:1000; Cell Signaling Technology, Inc., Danvers, MA), p53 (1:1000; cat. no. 10422-1-AP; ProteinTech Group, Inc., Rosemont, IL), p21 (1:1000; cat. no. 27296-1-AP; ProteinTech Group, Inc., Rosemont, IL), αSMA (1:1000, cat. no. sc-8432; Santa Cruz Biotechnology, Inc., Dallas, TX), fibronectin (1:1000, cat. no. sc-8422; Santa Cruz Biotechnology, Inc., Dallas, TX), Smad2 (1:1000; cat. no. sc-393312 Santa Cruz Biotechnology, Inc., Dallas, TX), Smad3 (1:1000; cat. no. sc-101154; Santa Cruz Biotechnology, Inc., Dallas, TX) or β-actin (1:1000; cat. no. sc-81178; Santa Cruz Biotechnology, Inc., Dallas, TX) at 4 °C overnight.

Techniques: Isolation, Expressing, Western Blot, Quantitative RT-PCR, Control, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Cell

Article Title: Contrasting somatic mutation patterns in aging human neurons and oligodendrocytes

doi: 10.1016/j.cell.2024.02.025

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Mouse monoclonal anti-SOX10 Alexa Fluor 647 , Novus Biologicals , Clone SOX10/991; catalog number: NBP2-59621.

Techniques: Whole Genome Amplification, Binding Assay, dsDNA Assay, Multiplex Assay, cDNA Synthesis, Sequencing, Software

Journal: bioRxiv

Article Title: Alteration of skin fibroblast steady state contributes to healing outcomes

doi: 10.1101/2024.12.06.627278

Figure Lengend Snippet: (A) Diagram of dermal fibroblast isolation and sequencing process; (B) Cell cluster UMAP plot; (B) Cell type distribution in each genotype; (D) Dot plot of selected marker genes for each cell cluster; (E) Feature plots of representative markers for Schwann cell development within Sox10+ population; (F) Histogram of Sox10-700 intensity of unstained and stained dermal fibroblasts and quantification of Sox10+ cells in WT and TSP2KO groups (n = 3, Unpaired T test, two-tailed, *, p<0.05).

Article Snippet: The following primary antibodies were used for flow cytometry, IHC or IF: Sox10 Antibody [Alexa Fluor 700] (1:200, Novus Biologics, NBP2-59621AF700), anti-Neurofilament H (NFH) (1:200, EMD Millipore, AB1989), anti-Vimentin (1:500, EMD Millipore, AB5733), TGFB3 Rabbit pAb (1:250, ABclonal, A8460), Sox10 Rabbit pAb (1:100, ABclonal, A15100), β-catenin (IF: 1:20, R&D system, AF1329).

Techniques: Isolation, Sequencing, Marker, Staining, Two Tailed Test

Journal: bioRxiv

Article Title: Alteration of skin fibroblast steady state contributes to healing outcomes

doi: 10.1101/2024.12.06.627278

Figure Lengend Snippet: (A) Immunofluorescence staining of NFH, Sox10, TGFb3 with Vimentin in D7 WT and TSP2KO mouse skin wounds. Quantification of (B) number of nerve bundles (NBs) in the center, (C) diameter of NBs (µm), (D) Sox10+ area/HPF (%), and (E) Tgfb3+ area/HPF (%) across the whole wounds (dashed circles indicate nerve bundles, n = 3-4, Unpaired T-test, two-tailed, *, p<0.05, **, p<0.01).

Article Snippet: The following primary antibodies were used for flow cytometry, IHC or IF: Sox10 Antibody [Alexa Fluor 700] (1:200, Novus Biologics, NBP2-59621AF700), anti-Neurofilament H (NFH) (1:200, EMD Millipore, AB1989), anti-Vimentin (1:500, EMD Millipore, AB5733), TGFB3 Rabbit pAb (1:250, ABclonal, A8460), Sox10 Rabbit pAb (1:100, ABclonal, A15100), β-catenin (IF: 1:20, R&D system, AF1329).

Techniques: Immunofluorescence, Staining, Two Tailed Test