Journal:

Article Title: The Chaperone Function of hsp70 Is Required for Protection against Stress-Induced Apoptosis

doi:

Figure Lengend Snippet: The C-terminal substitution mutant hsp70AAAA inhibits JNK activation without blocking its rate of dephosphorylation. IMR90 lung fibroblast cells (20 to 30 population doublings) were infected with an adenovirus expressing tTA (AdCMV-tTA) and an adenovirus encoding a tetracycline-regulated expression cassette encoding full-length hsp70 (+Hsp70) or hsp70 with the C-terminal four amino acids EEVD replaced with AAAA (+Hsp70-4AA). Control cells received the same total multiplicity of infection of the hsp70-encoding virus but without the Ad CMV-tTA virus (control). The infected cells were heat shocked for 30 min at 45°C and then incubated at 37°C with the protein kinase inhibitor staurosporine to block new JNK phosphorylation. Extracts were prepared from non-heat-shocked cells (Con.), heat-shocked cells (0), and heat-shocked cells that were incubated with staurosporine for 20 or 40 min at 37°C after the heat shock. Shown is an immunoblot of cytosolic extracts probed with an anti-phospho-JNK antibody.

Article Snippet: JNK activation was measured by Western blotting using a phosphorylation state-specific anti-JNK antibody (Promega Corp., Madison, Wis.).

Techniques: Mutagenesis, Activation Assay, Blocking Assay, De-Phosphorylation Assay, Infection, Expressing, Incubation, Western Blot

Journal: Scientific Reports

Article Title: Diacerein ameliorates cholestasis-induced liver fibrosis in rat via modulating HMGB1/RAGE/NF-κB/JNK pathway and endoplasmic reticulum stress

doi: 10.1038/s41598-023-38375-4

Figure Lengend Snippet: Bar charts showing the effect of oral administration of different doses of diacerein on ER stress-related parameters: ( A ) GRP78 gene expression (folds change), ( B ) IRE 1α gene expression (folds change), ( C ) PERK gene expression (folds change), and ( D ) p-JNK protein content (ng/g tissue) in liver tissue of different experimental groups. The values of different groups were presented as means ± SE (n = 8 rats per group). *p < 0.05 versus the sham-operated group; # p < 0.05 versus the BDL group; $ p < 0.05 versus the diacerein-treated group (10 mg/kg/day); + p < 0.05 versus the diacerein-treated group (30 mg/kg/day).

Article Snippet: The p-JNK ELISA kits purchased from RayBiotech (Life, Georgia, USA).

Techniques: Expressing

Journal: Nature Communications

Article Title: Mechanistic characterization of a Drosophila model of paraneoplastic nephrotic syndrome

doi: 10.1038/s41467-024-45493-8

Figure Lengend Snippet: A – D UMAP showing the gene expression changes of JNK/Jra pathway genes in MT of EGT > w1118 and EGT > yki act flies. E , F qPCR of Jra , kay and puc genes in MT of EGT > yki act and PC ts > Pvr act flies. Transgenes were induced for 8 days. Data are presented as means ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001 with student t-test. G – J Western blots showing the protein levels of JNK and phosphorylated JNK in MT of EGT > w1118 and EGT > yki act G and PC ts > w1118 and PC ts > Pvr act I flies. H shows the signal in the stem cell zone (SCZ) and principal cells plus stellate cells (PC + SC). J shows the rescue of p-JNK upon knockdown of Pvr in principal cells in yki act flies. Transgenes were induced for 5 and 8 days in G , I ; 8 days in H ; 4 and 7 days in J . n = 3 biologically independent experiments with 2 or 3 technical replicates in E , F .

Article Snippet: G – J Western blots showing the protein levels of JNK and phosphorylated JNK in MT of EGT > w1118 and EGT > yki act G and PC ts > w1118 and PC ts > Pvr act I flies.

Techniques: Gene Expression, Western Blot, Knockdown