Journal: Neural Regeneration Research

Article Title: Lithium promotes recovery after spinal cord injury

doi: 10.4103/1673-5374.327348

Figure Lengend Snippet: Lithium suppresses activation of inflammatory cytokines in the spinal cord of rats with SCI . Sham group: No spinal cord injury or treatment; SCI group: spinal cord injury only; LiCl group: lithium treatment after spinal cord injury. (A–C) TNF-α (A), IL-1β (B), and IL-6 (C) expression in the spinal cord were detected by enzyme-linked immunosorbent assay. (D) Representative immunohistochemical staining for IL-1β 7 days after surgery. The number of IL-1β-positive cells was higher in the LiCl group than in the sham group, but lower than in the SCI group. Black arrows indicate IL-1β-positive cells. Scale bars: 50 μm. (E) Quantification of IL-1β-positive cells 7 days after surgery. (F, G) TNF-α, IL-1β, and IL-6 expression. Data are shown as mean ± SD ( n = 6). * P < 0.05, vs . sham group; # P < 0.05, vs . SCI group (one-way analysis of variance followed by the least significant difference test). IL-1β: Interleukin-1β; IL-6: interleukin-6; LiCl: lithium chloride; SCI: spinal cord injury; TNF-α: tumor necrosis factor-alpha.

Article Snippet: Subsequently, the membranes were incubated overnight at 4°C with rat monoclonal primary antibodies against IL-6 (1:1000, Acris Antibodies GmbH, Herford, Germany; Cat# SM1695PX, RRID:AB_1004115), IL-1β (1:100, Miltenyi Biotec, Bergisch Gladbach, Germany; Cat# 130-125-220, RRID:AB_2889697), TNF-α (1:100, Miltenyi Biotec; Cat# 130-102-386, RRID:AB_2661141), NLRP3 (1:1000, MyBioSource, San Diego, CA, USA; Cat# MBS604215, RRID:AB_10911109), ASC (1:2000, Gabriel Núñez, University of Michigan, Ann Arbor, MI, USA; Cat# GN-ASC, RRID:AB_2750645), pro-Caspase-1 (1:1000, ProSci, San Diego, CA, USA; Cat# 48-495, RRID:AB_1945485), Caspase-1 (1:1000, Acris Antibodies GmbH; Cat# AP06610PU-N, RRID:AB_1611115), GSDMD (1:100, Thermo Fisher Scientific; Cat# A305-736A-M, RRID:AB_2782894), and IL-18 (1:100, DSHB, Iowa City, IA, USA; Cat# CPTC-IL18-2, RRID:AB_1553716), followed by incubation with horseradish peroxidase-conjugated rabbit anti-rat IgG (1:1000, Biorbyt, Cat# orb21564, RRID:AB_10932535) at 20°C for 1 hour, and finally stained with a diaminobenzidine solution.

Techniques: Activation Assay, Expressing, Enzyme-linked Immunosorbent Assay, Immunohistochemical staining, Staining

Journal: Neural Regeneration Research

Article Title: Lithium promotes recovery after spinal cord injury

doi: 10.4103/1673-5374.327348

Figure Lengend Snippet: Lithium attenuates inflammatory, oxidative, and pyroptotic injury in PC12 cells subjected to OGD . Con group: untreated PC12 cells; OGD group: PC12 cells subjected to OGD; OGD + LiCl group: PC12 cells subjected to OGD and treated with LiCl; OGD + LiCl + ATRA group: PC12 cells subjected to OGD and treated with LiCl and the Nrf2 inhibitor ATRA. (A–I) TNF-α (A), IL-1β (B), IL-6 (C), CAT (D), GSH-Px (E), LDH (F), SOD (G), MDA (H), and T-AOC (I) expression as detected by ELISA. (J–N) Flow cytometry analysis of the percentage of pyroptotic PC12 cells. Data are shown as mean ± SD ( n = 6). * P < 0.05, vs . control group; # P < 0.05, vs . OGD group; & P < 0.05, vs . OGD + LiCl group (one-way analysis of variance followed by least significant difference test). ATRA: All-trans retinoic acid; CAT: catalase; Con: control; GSH-Px: glutathione peroxidase; IL-1β: interleukin-1β; IL-6: interleukin-6; LiCl: lithium chloride; LPO: lipid peroxide; MDA: malondialdehyde; OGD: oxygen glucose deprivation; SOD: superoxide dismutase; T-AOC: total antioxidant capacity; TNF-α: tumor necrosis factor-alpha.

Article Snippet: Subsequently, the membranes were incubated overnight at 4°C with rat monoclonal primary antibodies against IL-6 (1:1000, Acris Antibodies GmbH, Herford, Germany; Cat# SM1695PX, RRID:AB_1004115), IL-1β (1:100, Miltenyi Biotec, Bergisch Gladbach, Germany; Cat# 130-125-220, RRID:AB_2889697), TNF-α (1:100, Miltenyi Biotec; Cat# 130-102-386, RRID:AB_2661141), NLRP3 (1:1000, MyBioSource, San Diego, CA, USA; Cat# MBS604215, RRID:AB_10911109), ASC (1:2000, Gabriel Núñez, University of Michigan, Ann Arbor, MI, USA; Cat# GN-ASC, RRID:AB_2750645), pro-Caspase-1 (1:1000, ProSci, San Diego, CA, USA; Cat# 48-495, RRID:AB_1945485), Caspase-1 (1:1000, Acris Antibodies GmbH; Cat# AP06610PU-N, RRID:AB_1611115), GSDMD (1:100, Thermo Fisher Scientific; Cat# A305-736A-M, RRID:AB_2782894), and IL-18 (1:100, DSHB, Iowa City, IA, USA; Cat# CPTC-IL18-2, RRID:AB_1553716), followed by incubation with horseradish peroxidase-conjugated rabbit anti-rat IgG (1:1000, Biorbyt, Cat# orb21564, RRID:AB_10932535) at 20°C for 1 hour, and finally stained with a diaminobenzidine solution.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Control

Journal: Bioengineered

Article Title: microRNA-128-3p inhibits CD4+ regulatory T cells enrichment by targeting interleukin 16 in gastric cancer

doi: 10.1080/21655979.2021.2017566

Figure Lengend Snippet: IL16 expression inversely correlated with miR-128-3p expression and was positively related to CD4+ Tregs infiltration in GC. (a) Venn diagram: The quartile distribution of miR-128-3p expression was used to divide GC tissues into low, medium and high expression groups. Three subsets, namely, upregulated genes in the low miR-128-3p group, cytokine-related genes, and genes with potential miR-128-3p binding sites, intersected. (b) Heatmap of IL16, SEMA4C, and STC1 expression in the low and high miR-128-3p groups. The color intensity corresponds to the enrichment Z-score: blue represents low expression, and red represents high expression. (c) Different scores for the expression of IL16, SEMA4C, and STC1 in the low miR-128-3p group. (d) Correlation analysis of miR-128-3p expression and IL-16 mRNA expression in 368 GC clinical samples in TCGA database. (e) Heatmap of the expression levels of IL16, enrichment score of CD4 + T cells and expression levels of miR-128-3p in 402 TCGA GC samples. Color intensity corresponds to the expression level Z-score: blue represents low expression, and red or yellow represents high expression. (f-h) Correlations between IL-16 and miR-128-3p (f), CD4 (g) or Foxp3 (h) in GC. (i) The expression of CD4, IL16 and Foxp3 and miR-128-3p mRNA levels was assessed by Western blotting and RT–qPCR. T: GC tissues; N: adjacent noncancerous gastric tissues. (j) The protein expression level of CD4, IL16 and Foxp3 was detected by immunohistochemistry.

Article Snippet: Paraffin sections were immunostained with antibodies against IL16 (1:100; Boster Biological Technology Co., Ltd., Beijing, China; PB9238), CD4 (1:200; bs-0647 R) and Foxp3 (1:500; ab20034) overnight at 4°C.

Techniques: Expressing, Binding Assay, Western Blot, Quantitative RT-PCR, Immunohistochemistry

Journal: Bioengineered

Article Title: microRNA-128-3p inhibits CD4+ regulatory T cells enrichment by targeting interleukin 16 in gastric cancer

doi: 10.1080/21655979.2021.2017566

Figure Lengend Snippet: IL16 was a target of miR-128-3p. (a, b) The efficiency of miR-128-3p overexpression or knockdown was examined using RT–qPCR. The expression of IL16 was measured in miR-128-3p overexpression group, miR-128-3p-knockdown group and the corresponding control group by Western blotting (c) and ELISA (d, e). (f) Schematic diagram of the predicted binding sites between miR-128-3p and IL16 WT 1 or 2 and the mutated binding region of IL16 MUT 1 or 2. (g, h) The interaction between miR-128-3p and IL16 was verified by dual-luciferase reporter assay. *P < 0.05, **P < 0.01. GC, gastric cancer; WT, wild type; MUT, mutant; NC, negative control.

Article Snippet: Paraffin sections were immunostained with antibodies against IL16 (1:100; Boster Biological Technology Co., Ltd., Beijing, China; PB9238), CD4 (1:200; bs-0647 R) and Foxp3 (1:500; ab20034) overnight at 4°C.

Techniques: Over Expression, Knockdown, Quantitative RT-PCR, Expressing, Control, Western Blot, Enzyme-linked Immunosorbent Assay, Binding Assay, Luciferase, Reporter Assay, Mutagenesis, Negative Control

Journal: Bioengineered

Article Title: microRNA-128-3p inhibits CD4+ regulatory T cells enrichment by targeting interleukin 16 in gastric cancer

doi: 10.1080/21655979.2021.2017566

Figure Lengend Snippet: IL16 promotes the differentiation of CD4+ CD25+ Foxp3+ Treg cells. T lymphocytes were cultured with RPMI medium containing recombinant IL16 (a) or IL10 (b), medium containing recombinant IL16 (c) or recombinant IL16 and anti-CD4 BA (d), or conditioned medium treated with LV-miR-128-3p-inhibitor (e) or treated with LV-miR-128-3p-inhibitor-containing anti-CD4 BA (f) for 48 h, and then, the populations of CD4+ CD25+ Foxp3+ Tregs were analyzed by flow cytometry. (g) A schematic of the molecular mechanism by which miR-128-3p affects the infiltration of CD4+ Tregs *P < 0.05, **P < 0.01. anti-CD4 BA, anti-CD4 was blocking antibody.

Article Snippet: Paraffin sections were immunostained with antibodies against IL16 (1:100; Boster Biological Technology Co., Ltd., Beijing, China; PB9238), CD4 (1:200; bs-0647 R) and Foxp3 (1:500; ab20034) overnight at 4°C.

Techniques: Cell Culture, Recombinant, Flow Cytometry, Blocking Assay