Journal: Journal of Functional Foods

Article Title: Abrus cantoniensis Hance alleviates APAP-triggered acute liver damage via inhibiting the MAPK and NF-κB signaling pathways in mice

doi: 10.1016/j.jff.2024.106145

Figure Lengend Snippet: Fig. 4. AC attenuated the hepatocytes apoptosis via inhibiting MAPK signaling pathway. (A) The level of JNK/p-JNK and p38/p-p38 were investigated by WB. (B-C) Quantification of liver tissue p-JNK/p-P38 protein level. (D-F) the effects of AC and JNK specific inhibitor SP600125 on Bcl-2/Bax expression in APAP-induced HepG2 cell. (G-I) the effects of AC on Bcl-2/Bax expression after the knockdown of JNK via specific siRNA. (J-L) the effects of AC and p38 MAPK specific inhibi tor SB203580 on Bcl-2/Bax expression in APAP-induced HepG2 cell. (M−O) the effects of AC on Bcl-2/Bax expression after the knockdown of p38 MAPK via specific siRNA. n = 3. Value was shown as mean ± Standard deviation (SD). Compared with the normal group, #P < 0.05, ##P < 0.01, ###P < 0.001; Compared with the model group, *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: The JNK1 siRNA (sc-29380), P38α MAPK siRNA (sc-29433), NFκB p65 siRNA (sc-29410) and scrambled siRNA were purchased from Santa Cruz Biotechnology.

Techniques: Expressing, Knockdown, Standard Deviation

Journal: Journal of Functional Foods

Article Title: Abrus cantoniensis Hance alleviates APAP-triggered acute liver damage via inhibiting the MAPK and NF-κB signaling pathways in mice

doi: 10.1016/j.jff.2024.106145

Figure Lengend Snippet: Fig. 6. AC alleviated the inflammation response by inhibiting NF-κB signaling pathway. (A) Liver tissue IκB α/p-IκB α protein content was determined by WB. (B) Cytosolic and nuclear p65 proteins was detected by WB. (C-E) Quantification of interest protein expression in liver tissue. (F-G) the effects of AC and NF-κB specific inhibitor PDTC on IL-1β and TNF-α production in LPS-induced THP-1 cell. (H-J) the effects of AC and PDTC on iNOS expression in LPS-induced THP-1 cell. (K-M) the effects of AC on iNOS expression after the knockdown of NF-κB p65 via specific siRNA. n = 3. Value was shown as mean ± Standard deviation (SD). Compared with the normal group, #P < 0.05, ##P < 0.01, ###P < 0.001; Compared with the model group, *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: The JNK1 siRNA (sc-29380), P38α MAPK siRNA (sc-29433), NFκB p65 siRNA (sc-29410) and scrambled siRNA were purchased from Santa Cruz Biotechnology.

Techniques: Expressing, Knockdown, Standard Deviation

Journal: International Journal of Biological Sciences

Article Title: Antrodia cinnamomea Galactomannan Elicits Immuno-stimulatory Activity Through Toll-like Receptor 4

doi: 10.7150/ijbs.24564

Figure Lengend Snippet: ACP induces cytokine expression through MAPK. ( A ) J774A.1 macrophages were incubated for 0-60 min with or without ACP (100 µg/mL). The phosphorylation levels of ERK1/2, JNK1/2 and p38 were assayed by Western blot. ( B, C ) J774A.1 macrophages stably transfected with a control shRNA plasmid (sh-SC), a ERK1 shRNA plasmid (sh-ERK1), a JNK1 shRNA plasmid (sh-TLR2), a p38α shRNA plasmid (sh-p38α), and a p38β shRNA plasmid (sh-p38β) were incubated for 24 h with or without ACP (100 μg/mL). The levels of TNF-α ( B ) and IL-6 ( C ) in the culture medium were measured by ELISA. The Western blot results are representative of those obtained in three different experiments. The ELISA data are expressed as the means ± SD of three separate experiments. *** indicates a significant difference at the level of p < 0.001.

Article Snippet: All plasmids were purchased from Santa Cruz Biotechnology (Santa Cruz, CA): ERK1 shRNA plasmid (sh-ERK1, sc-29308-SH), JNK1 shRNA plasmid (sh-JNK1, sc-29381-SH), p38α shRNA plasmid (sh-p38α, sc-29434-SH), p38β shRNA plasmid (sh-p38β, sc-39117-SH), TLR2 shRNA plasmid (sh-TLR2, sc-40257-SH) and TLR4 shRNA plasmid (sh-TLR4, sc-40261-SH).

Techniques: Expressing, Incubation, Western Blot, Stable Transfection, Transfection, shRNA, Plasmid Preparation, Enzyme-linked Immunosorbent Assay

Journal: Scientific reports

Article Title: Up-regulation of ST18 in pemphigus vulgaris drives a self-amplifying p53-dependent pathomechanism resulting in decreased desmoglein 3 expression.

doi: 10.1038/s41598-022-09951-x

Figure Lengend Snippet: Figure 2. Antibody-mediated DSG3 down-regulation affects p53 expression and activity in a p38MAPK- dependent manner. (a) NHEKs exposed to either AK23 or negative control antibody (NC) were stained for p53 (green) and DAPI (blue) (scale bar = 20um); (b) p53 protein expression in NHEKs exposed to either AK23 or negative control antibody (NC) was assessed using immunoblotting with anti-p53 antibody. β-actin served as a loading control (left panel). Protein levels were quantified and data was normalized to levels observed in negative control antibody-treated cells (right panel). Results represent the mean ± SE of four independent experiments (*p < 0.05 by 2-tailed t test); (c) NHEKs were transfected with a luciferase reporter construct under the regulation of a p53 binding site or with a control reporter, and were then treated either with AK23 antibody or negative control antibody (NC). Results represent the mean ± SE of three independent experiments (***p < 0.001 by 2-tailed t test); (d) NHEKs were transfected with a luciferase reporter construct under the regulation of a p53 binding site or a control reporter. Cells were additionally transfected with control (si-control) or p38MAPK-specific (si-p38) siRNAs. Twenty four hours post-transfection, cells were treated either with AK23 antibody (AK23 Ab) or negative control antibody (NC Ab). Results represent the mean ± SE of four independent experiments (*p < 0.05 by one way ANOVA test). Original blots are presented in Supplementary Fig. 4.

Article Snippet: Briefly, primary keratinocytes were co-transfected with the same pGL2 vectors, Renilla expression vector and control siRNA (Life Technologies, Carlsbad, CA, 452002) or a specific p38 alpha MAPK14-specific siRNA (Santa Cruz Biotechnology, Santa Cruz, CA, SC-29433).

Techniques: Expressing, Activity Assay, Negative Control, Staining, Western Blot, Control, Transfection, Luciferase, Construct, Binding Assay

Journal: Scientific reports

Article Title: Up-regulation of ST18 in pemphigus vulgaris drives a self-amplifying p53-dependent pathomechanism resulting in decreased desmoglein 3 expression.

doi: 10.1038/s41598-022-09951-x

Figure Lengend Snippet: Figure 4. ST18 drives a p53-dependent self-amplifying process promoting autoantibody-mediated membranal DSG3 down-regulation in pemphigus vulgaris. Genetically determined ST18 overexpression (1) enhances autoantibodies-induced DSG3 down-regulation in keratinocytes (2, orange arrow) which triggers p38MAPK- dependent (3) p53 activity (4), which in turn up-regulates ST18 promoter activity (5), thus setting the stage for a self-amplifying pathogenetic cycle in PV.

Article Snippet: Briefly, primary keratinocytes were co-transfected with the same pGL2 vectors, Renilla expression vector and control siRNA (Life Technologies, Carlsbad, CA, 452002) or a specific p38 alpha MAPK14-specific siRNA (Santa Cruz Biotechnology, Santa Cruz, CA, SC-29433).

Techniques: Over Expression, Activity Assay