Journal: Oncogenesis

Article Title: Post-transcriptional screen of cancer amplified genes identifies ERBB2 / Her2 signaling as AU-rich mRNA stability-promoting pathway

doi: 10.1038/s41389-021-00351-w

Figure Lengend Snippet: A Immunoblot analysis of ERBB2, AKT, MEK1/2, and ERK1/2 signaling inhibition after treatment with increasing doses of lapatinib for 24 h. Status of ERBB2 (phospho-tyrosine 1221/1222 and total), AKT (phospho-serine 473 and total pan AKT), ERK1/2 (Phospho-threonine 202/phospho-tyrosine 204 and total), MEK1/2 (Ser217/221), and ZFP36/TTP are shown. A blot from at least two experiments is shown. B Immunoblot analysis for the inhibition of MK2 activation after treatment with lapatinib. SKBR3 cells were treated with increasing doses of lapatinib, as indicated for 24 h. Phospho-threonine MK2 (T222) and total MK2 protein abundance are shown. A representative blot from two experiments is shown. C Immunoblot analysis for ZFP36/TTP phosphorylation in MAPKAPK-2 silenced SKBR3 cells. SKBR3 cells transfected with 100 nM of si Ctrl or si MAPKAPK-2 for 24 h were tested for ZFP36/TTP protein abundance. Total MK2, phosphorylated ZFP36/TTP, and β-actin protein abundances are shown. A representative blot from three experiments is shown. Effect of the MK2 inhibitor PF-3644022 (1 µM, 4 h) on the abundance of phosphorylated ZFP36/TTP in ERBB2-expressing SKBR3 ( D ) or SKBR3 transfected with ZFP36 expression plasmid ( E ). Western blots are from two independent experiments. F Immunoblot analysis of total and phosphorylated ZFP36/TTP, and MK2 in MK2 -knockout MEF cells in the absence or presence of overnight transfected ERBB2.

Article Snippet: PF-3644022 (MK2 inhibitor) was purchased from Tocris (Tocris Bioscience, Minneapolis, MN).

Techniques: Western Blot, Inhibition, Activation Assay, Transfection, Expressing, Plasmid Preparation, Knock-Out

Journal: iScience

Article Title: NLRP6 induces RIP1 kinase-dependent necroptosis via TAK1-mediated p38 MAPK /MK2 phosphorylation in S . typhimurium infection

doi: 10.1016/j.isci.2024.109339

Figure Lengend Snippet: NLRP6 promotes the systemic dissemination of S . typhimurium by regulating MK2/RIP1-associated necroptosis in vivo (A–F) Streptomycin-pretreated WT and Nlrp6 −/− mice were orally infected with 1 × 10 8 CFU of S . typhimurium strain SL1344. Mice were administrated intraperitoneally with the RIP1 inhibitor Nec-1s at 1 h prior to infection and 24 h post-infection, followed by sample collection at 2 days post-infection. Western blot analysis and relative intensity of colonic intestinal epithelial cells isolated from mice (A, B). Bacterial burden in MLN (C), liver (D) and spleen (E). Intestinal permeability (F). (G and H) Streptomycin-pretreated WT and Nlrp6 −/− mice were orally infected with 1 × 10 8 CFU of SL1344 and analyzed at 2 days post-infection. Western blot analysis and relative intensity of colonic intestinal epithelial cells isolated from mice. (I–M) Streptomycin-pretreated WT and Nlrp6 −/− mice were orally infected with 1 × 10 8 CFU of SL1344 and administrated orally with the MK2 inhibitor PF-3644022 at 4 h post-infection, followed by sample collection at 2 days post-infection. Western blot analysis and relative intensity of colonic intestinal epithelial cells isolated from mice (I, J). Bacterial burden in MLN (K), liver (L) and spleen (M). Data were compared with independent Student’s t test. Values are expressed as the mean ± SEM, and statistically significant differences are indicated. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ns, not significant.

Article Snippet: PF-3644022 , Selleck Chemicals , Cat# S8224.

Techniques: In Vivo, Infection, Western Blot, Isolation, Permeability