p38 sirna santa cruz biotechnology  (Santa Cruz Biotechnology)

Journal: Nature Communications

Article Title: Stress pathway outputs are encoded by pH-dependent clustering of kinase components

doi: 10.1038/s41467-024-50638-w

Figure Lengend Snippet: A HEK293 cells, depleted of JNK1 with siRNA (1 to 25 pmol, n = 4) or treated with control siRNA (25 pmol, n = 12), were stimulated with TNFα (10 ng/ml) and change (Δ) in JNK activity (KTR) and pHi monitored over time (60 min). B TNFα-stimulated change in JNK activity and pHi in HEK293 cells depleted of JNK2 ( n = 3, 4, 3 for 1, 6, 25 pmol). C TNFα-stimulated change in pHi was correlated with JNK activity responses in JNK1 or JNK2-depleted cells. D HEK293 cells, transfected with JNK1 ( n = 4, 3, 3 for 1, 6, 25 pmol) or control siRNA (25 pmol, n = 12), were stimulated with sorbitol (150 mM) and JNK activity and pHi change measured over time. E Sorbitol stimulated change in JNK activity and pHi in HEK293 cells depleted of JNK2 ( n = 4, 4, 3 for 1, 6, 25 pmol). F Sorbitol-stimulated change in pHi was correlated with JNK activity responses in JNK1 or JNK2-depleted cells. Values represent mean ± SEM from independent experiments (n). ΔKTR and ΔpHi values indicate a difference compared to before stimulation (0 min). ‘R’ indicates Pearson correlation coefficients between stress-induced change in JNK activity and pHi.

Article Snippet: For siRNA-mediated JNK1 and JNK2 knockdown, cells were transfected with 1, 6 or 25 pmol ON-TARGET plus Human MAPK8 SMART pool siRNA (Dharmacon, L-003514-00-0020) and/or ON-TARGET plus Human MAPK9 SMART pool siRNA (Dharmacon, L-003505-00-0020) with 2 µl RNAiMAX (Thermo Fisher Scientific), according to the manufacturer’s protocol.

Techniques: Control, Activity Assay, Transfection

Journal: Cell Death Discovery

Article Title: Interleukin-1β induces and accelerates human endometrial stromal cell senescence and impairs decidualization via the c-Jun N-terminal kinase pathway

doi: 10.1038/s41420-024-02048-6

Figure Lengend Snippet: A ELISA confirmed that CCL5 secretion by P3 ESC from each of four subjects was upregulated after 72 h of IL-1β treatment. This effect was reversed by co-incubation with SP ( p < 0.001, ANOVA). Error bars indicate SD of triplicate determinations for subject S4. B When IL-1β was co-incubated with a 100-fold excess of IL-1ra , complete abrogation of the IL-1β effect on IL-8 and IL-6 upregulation was observed. The overall comparison using two-factor ANOVA analysis revealed significant differences ( p < 0.001). Scheffé Test comparisons showed significant differences between Control vs IL-1β (p < 0.001) and IL-1β vs IL-1ra ( p < 0.001). Error bars indicate SD of triplicate determinations. C Time-course of CCL5 ELISA response to IL-1β ± SP (24–72 h). The overall comparison using one-factor ANOVA revealed significant differences between groups (p < 0.001). Error bars indicate SD of triplicate determinations. D Time-course of TNF-α ELISA response to IL-1β ± SP (24–72 h). The overall comparison using one-factor ANOVA revealed significant differences between groups ( p < 0.001). Error bars indicate SD of triplicate determinations. E Western blot showing that preincubation of P3 ESC with SP, for 1 h before addition of IL-1β (PSP), had the same inhibitory effect as simultaneous co-incubation of IL-1β plus SP (SSP) on IL-1β, IL-6, TNF-α and MMP3 reduction. SP alone reduced basal (control) SASP markers. β-Actin levels were not affected. F Western blot reveals inhibition of IL-1β stimulation of IL-1β, IL - 6, MMP3, p21 and HMGB1 by JNK1 (CJ1) or JNK2 (CJ2) siRNA constructs transfected into P3 ESC. β-Actin levels were not affected.

Article Snippet: As an alternative approach to block JNK action in ESC, JNK1 and JNK2 double-stranded siRNA constructs or a scrambled duplex control purchased from Cell Signaling Technologies (cat# 6232, 6233 and 6568), were introduced into ESC via transient transfection at a concentration of 100 nmol per well using Lipofectamine RNAiMAX (Thermo-Fisher) as described [ ].

Techniques: Enzyme-linked Immunosorbent Assay, Incubation, Comparison, Western Blot, Inhibition, Construct, Transfection