il 1β  (Santa Cruz Biotechnology)

Journal: Brain, behavior, and immunity

Article Title: The role of the Toll like receptor 4 signaling in sex-specific persistency of depression-like behavior in response to chronic stress

doi: 10.1016/j.bbi.2023.10.006

Figure Lengend Snippet: (A) Experimental outline. Unstressed mice that were not exposed to chronic stress and age-matched (upper panel) and stressed mice that were exposed to chronic stress exposure for 28 days, followed by 28 days of post-stress period. (B and C) Bar graphs showing total immobility time in female (red bar) and male (blue bar) stressed mice compared to unstressed mice at D28 (B) and at D56(C). Statistical analyses were performed using Two-way ANOVA (stress effect, p < 0.0001; sex effect, p = 0.2065; interaction effect, p = 0.0030; * p < 0.05, **** p < 0.001 compared to unstressed male mice; #### p < 0.0001, compared to unstressed female mice, + p < 0.05, +++ p < 0.001, compared to stressed male mice, n = 7–9 mice for each group). (D) A graph showing depression-like behavior over the entire experimental, with female mice shown in red and male mice shown in blue. (E-N) Microglia activation and morphological analysis in female and male mice at D56. (E and J) Representative immunofluorescence images of IBA-1 + (red) and CD68 + (yellow) cells in female (E.1 and E.2) and male (J.1 and J.2) mice. (F and K) Bar graphs showing number of IBA-1 + cells in female (F) and male (K) mice. (G and L) Bar graphs showing quantification of CD68 + area per IBA-1 + microglia in female (G) and male (L) mice. (E and J) Representative images of three-dimensional (3D) reconstruction in female (E.3, E.4, E.5 and E.6) male (J.3, J.4, J.5 and J.6) mice. The outlined with a white box is magnified. (H-N) Sholl analysis based on 3D reconstruction in female and male mice. Bar graph showing soma size per microglia in female (H) and male (M) mice. Line graphs showing quantification of number of intersection between each circle (green, E5, E6, J5 and J6) in female (I) and male (N) mice. Statistical analyses were performed using t -test (* p < 0.05, compared to unstressed female mice). Scale bars indicate 20 (E and J.1–2) and 10 (E and J.3–6) um. Each dot represents an individual mouse. Results are expressed as the mean ± SEM.

Article Snippet: The membrane was then blocked for 1 h at room temperature and treated overnight with primary antibodies specific to p-NF-kB p65 (3033 s, Abcam), NF-kB p65 (51–0500, Thermo Fisher Scientific), caspase-1 (20B-0042, adipogen, CA, USA), IL-1β (6243S, Cell signaling, MA, USA), and α-tubulin (T9026, Sigma) at 4 °C.

Techniques: Activation Assay, Immunofluorescence

Journal: Brain, behavior, and immunity

Article Title: The role of the Toll like receptor 4 signaling in sex-specific persistency of depression-like behavior in response to chronic stress

doi: 10.1016/j.bbi.2023.10.006

Figure Lengend Snippet: (A) The experimental outline in Cx3Cr1 CreEr ; Tlr4 fl double transgenic mice. Unstressed female mice that were not exposed to chronic stress and age-matched and stressed female mice that were exposed to chronic stress for 28 days, followed by 28 days of post-stress period, and then treated with vehicle or tamoxifen for an additional 12 days. (B-D) Bar graphs showing total immobility time at D28 (B), D56 (C) and D68 (D). (E-G) Representative western blot of p-NF-KB 65 (E, upper panel), NF-kB p65 (E, upper panel), caspase-1 (F, upper panel) and IL-1β (G, upper panel) and bar graphs showing relative quantification of p-NF-KB p65 (E, bottom left panel) normalized to NF-kB p65, NF-kB p65 (E, bottom right panel), cleaved caspase-1 (F, bottom panel) and IL-1β (G, bottom panel) levels normalized to α-tubulin. (H) Bar graph showing concentration of IL-1β in PFC lysates. Statistical analyses were performed using t -test or One-way ANOVA (* p < 0.05, ** p < 0.01, **** p < 0.0001 compared to female unstressed mice; # p < 0.05, ## p < 0.01, ### p < 0.001, compared to female stressed mice, n = 4–5 mice for each group). Each dot represents an individual mouse. Results are expressed as the mean ± SEM.

Article Snippet: The membrane was then blocked for 1 h at room temperature and treated overnight with primary antibodies specific to p-NF-kB p65 (3033 s, Abcam), NF-kB p65 (51–0500, Thermo Fisher Scientific), caspase-1 (20B-0042, adipogen, CA, USA), IL-1β (6243S, Cell signaling, MA, USA), and α-tubulin (T9026, Sigma) at 4 °C.

Techniques: Transgenic Assay, Western Blot, Quantitative Proteomics, Concentration Assay

Journal: Brain, behavior, and immunity

Article Title: The role of the Toll like receptor 4 signaling in sex-specific persistency of depression-like behavior in response to chronic stress

doi: 10.1016/j.bbi.2023.10.006

Figure Lengend Snippet: (A) Schematic showing TRAP-sequencing. (B) Selected Gene ontology (GO) annotations enriched in GFP-bound (green, upper panel) and GFP-unbound group (gray, lower panel).The x-axis represents - log10 (p-value), with dotted line representing a p -value of 0.05. (C-D) Volcano plots in female (C) and male (D) mice. The x-axis represents the log 2 conversion of the fold change (log 2 FC) values, and the y-axis represents the corrected significance level after base log10 conversion ( p value). Red and blue dots in the volcano plot indicate all DEGs that were found to differ significantly (absolute value of log 2 FC > 1, * p < 0.05). (E) Heat map with hierarchical clustering distances shows the variation in the expression levels (VST-scored log 2 RPKM) between female stressed and unstressed mice. (F) A bar graph showing selected GO annotations enriched in female (red) and male (blue) mice. The x-axis represents -log10 ( p -value), with dotted line representing a p -value of 0.05. (G) A bar graph showing log 2 FC of significant DEGs in female stress (red) and male stress (blue) mice using a cut-off of p < 0.05.

Article Snippet: The membrane was then blocked for 1 h at room temperature and treated overnight with primary antibodies specific to p-NF-kB p65 (3033 s, Abcam), NF-kB p65 (51–0500, Thermo Fisher Scientific), caspase-1 (20B-0042, adipogen, CA, USA), IL-1β (6243S, Cell signaling, MA, USA), and α-tubulin (T9026, Sigma) at 4 °C.

Techniques: Sequencing, Expressing

Journal: Cardiovascular Diagnosis and Therapy

Article Title: MicroRNA-155 inhibition attenuates myocardial infarction-induced connexin 43 degradation in cardiomyocytes by reducing pro-inflammatory macrophage activation

doi: 10.21037/cdt-21-743

Figure Lengend Snippet: Effects of miR-155 inhibition on MI- and LPS-induced IL-1β and MMP7 expression in vivo and in vitro. (A,B) Representative Western blots of IL-1β and MMP7 expression in cardiac tissue following MI. (C,D) Quantitative analysis of data in (A) and (B), respectively (n=5; *, P<0.01 vs. the sham group; $, P<0.05 vs. the MI group). (E,F) Representative Western blots of IL-1β and MMP7 expression in LPS-induced BMDMs with or without miR-155 antagomir treatment for 24 h. (G,H) Quantitative analysis of data in (E) and (F), respectively (n=3; *, P<0.01 vs. the control group; $, P<0.05 vs. the LPS group). Con, control; MI, myocardial infarction; LPS, lipopolysaccharide; BMDMs, bone-marrow-derived macrophages; IL-1β, interleukin-1β; MMP7, matrix metalloproteinase 7.

Article Snippet: The control con/miR-155-agomir, con/miR-155-antagomir (scrambled sequence), miR-155 antagomir (a 2'-O-methyl + 5' cholesterol-modified miR-155 inhibitor), small interfering (si)RNA control, and siRNAs specific for SOCS1, IL-1β, and MMP7 were purchased from Guangzhou RiboBio Co., Ltd. (Guangzhou, China).

Techniques: Inhibition, Expressing, In Vivo, In Vitro, Western Blot, Control, Derivative Assay

Journal: Cardiovascular Diagnosis and Therapy

Article Title: MicroRNA-155 inhibition attenuates myocardial infarction-induced connexin 43 degradation in cardiomyocytes by reducing pro-inflammatory macrophage activation

doi: 10.21037/cdt-21-743

Figure Lengend Snippet: Effect of miR-155 inhibition on Cx43 degradation in vivo post-MI and in vitro in hypoxic NRCMs. (A) Representative photomicrographs of Cx43 expression (red staining) in mouse cardiac tissue after MI. Nuclei were counterstained with DAPI. Scale bar, 20 µm. (B) Representative Western blots of Cx43 expression in mouse cardiac tissue after MI. (C) Quantitative analysis of data in (B) (n=5; *, P<0.01 vs. the sham group; $, P<0.05 vs. the MI group). (D-I) Conditioned medium was obtained from LPS-induced BMDMs treated with or without miR-155 antagomir/agomir for 24 h and then applied to cultures of hypoxia-induced NRCMs for 24 h. (D,F,H) Representative Western blots of Cx43 expression. (E,G,I) Quantitative densitometric analysis of data from (D), (F) and (H), respectively [n=3; *, P<0.05 vs. the Con, the hypoxia + medium (LPS + con-antagomir/con-agomir) or the hypoxia + medium (LPS + miR-155 con-agomir) group]. (J-Q) LPS-induced BMDMs were treated with or without si-IL-1β RNA, si-MMP7 RNA or miR-155 agomir for 24 h, and the resulting conditioned medium was applied to hypoxia-induced NRCMs for 24 h. (J,L,N,P) Representative Western blots of IL-1β, MMP7 and Cx43 expression. (K,M,O,Q) Quantitative densitometric analysis of data from (J), (L), (N) and (P), respectively [n=3; *, P<0.05 vs. the si-con or the hypoxia + medium (LPS + con-agomir) group; $, P<0.05 vs. the hypoxia + medium (LPS + si-IL-1β or LPS + si-MMP7) group]. Con, control; Cx43, connexin 43; MI, myocardial infarction; LPS, lipopolysaccharide; BMDMs, bone-marrow-derived macrophages; NRCMs, neonatal mouse cardiomyocytes; IL-1β, interleukin-1β; MMP7, matrix metalloproteinase 7.

Article Snippet: The control con/miR-155-agomir, con/miR-155-antagomir (scrambled sequence), miR-155 antagomir (a 2'-O-methyl + 5' cholesterol-modified miR-155 inhibitor), small interfering (si)RNA control, and siRNAs specific for SOCS1, IL-1β, and MMP7 were purchased from Guangzhou RiboBio Co., Ltd. (Guangzhou, China).

Techniques: Inhibition, In Vivo, In Vitro, Expressing, Staining, Western Blot, Control, Derivative Assay

Journal: Cardiovascular Diagnosis and Therapy

Article Title: MicroRNA-155 inhibition attenuates myocardial infarction-induced connexin 43 degradation in cardiomyocytes by reducing pro-inflammatory macrophage activation

doi: 10.21037/cdt-21-743

Figure Lengend Snippet: Summary of the roles of miR-155 in the MI-induced connexin 43 degradation in cardiomyocytes by activating pro-inflammatory macrophage activation. MI, myocardial infarction; LPS, lipopolysaccharide; SOCS1, suppressor of cytokine signaling 1; IL-1β, interleukin-1β; MMP7, matrix metalloproteinase 7; NF-κB, nuclear factor κB.

Article Snippet: The control con/miR-155-agomir, con/miR-155-antagomir (scrambled sequence), miR-155 antagomir (a 2'-O-methyl + 5' cholesterol-modified miR-155 inhibitor), small interfering (si)RNA control, and siRNAs specific for SOCS1, IL-1β, and MMP7 were purchased from Guangzhou RiboBio Co., Ltd. (Guangzhou, China).

Techniques: Activation Assay