Journal: bioRxiv

Article Title: Fasting reverses drug-resistance in hepatocellular carcinoma through p53-dependent metabolic synergism

doi: 10.1101/2021.02.10.430545

Figure Lengend Snippet: (A) Viability of HepG2 cells after 24 h of incubation in growth medium (GM) or starvation medium (SM) with indicated concentrations of doxorubicin (Doxo). Values are normalized to the viability of non-treated cells grown in GM. (B) Viability of HepG2 cells after 24 h of incubation in GM or SM with indicated concentrations of Doxo and/or sorafenib (Sfb). Values are normalized to the viability of non-treated cells grown in GM. Comparison of groups treated with Doxo and Sfb vs corresponding groups treated with Sfb is shown. (C) HepG2 cells were grown in GM or SM with 10 μM sorafenib for indicated times, and western blot analysis was performed to determine protein levels of AKT, ERK, 4E-BP1, S6K and their respective phosphorylated forms. GAPDH served as loading control. (D) Western blot analysis was performed to determine protein levels of AKT, ERK, 4E-BP1, S6K and their respective phosphorylated forms in HepG2 xenografts from ad libitum fed (FED) and intermittently fasted (IF) nude mice treated with 30 mg/kg Sfb. GAPDH served as loading control. (E) Quantification of phosphorylated protein levels, relative to GAPDH and normalized to the protein level in xenograft protein lysates from FED mice. Unpaired t-test was performed. (F) Viability of HepG2 cells after 24 h of incubation GM or SM with Sfb and/or MEK inhibitor (5 µM U0126), ERK1/2 inhibitor (1 µM SCH772984), PI3K inhibitor (1 µM Wortmannin), PI3K inhibitor (1 µM MK-2206), 10 nM Rapamycin and 10 nM Bafilomycin determined. Comparison of groups within each inhibitor treatment group are shown. If not noted otherwise, mean values ± SEM are shown and two-way ANOVA, Tukey’s multiple comparisons test was performed. *** p<0.001, ** p<0.01, * p<0.05, ns-not significant (p>0.05); O.D. optical density.

Article Snippet: Following inhibitors were used: SCH 772984 ERK1/2 inhibitor (Cayman Chemicals), U0126 MEK inhibitor (Promega), Wortmannin (HY-10197-10mM/1mL), Buparlisib (HY-70063-10mM/1mL) and MK-2206 dihydrochloride (HY-10358-10mM/1mL) all from MedChemExpress, Bafilomycin A1 (BioMol), Rapamycin (Life Technologies, NOVEX).

Techniques: Incubation, Western Blot

Journal: Poultry Science

Article Title: GH inhibits ALV-J replication and restricts cell cycle by activating PI3K/Akt signaling pathway ☆

doi: 10.1016/j.psj.2024.104514

Figure Lengend Snippet: GH inhibited the cell cycle by activating PI3K/Akt pathway during ALV-J infection. DF-1 cells were transfected with GH overexpression vector (pcDNA3.1-GH) or GH knockdown vector (si-GH) in DF-1 cells for 24 h and then infected with ALV-J (TCID 50 = 10 4 ) for 12 and 24 h (pcDNA3.1 and si-NC served as negative controls). (A, B) Immunoblot analysis of the expression of env, Akt, p-Akt, and mTOR for each indicated time (β actin was used as loading control). (C, D) Cells were treated with p38, ERK, or PI3K phosphorylation inhibitors for 6 h following GH overexpression for 24 h, and then infected with ALV-J (TCID 50 = 10 4 ) for 24 h. Immunoblot analysis of env expression was performed. (E-H) qRT-PCR analysis of the relative mRNA levels of genes involved in cell proliferation (E, G) and apoptosis (F, H). P values were calculated using two-tail unpaired Student’s t-test. ( p < 0.05). * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: The p38 MAPK inhibitor (SB203580) and ERK1/2 inhibitor (SCH772984) were purchased from Selleck (Shanghai, China).

Techniques: Infection, Transfection, Over Expression, Plasmid Preparation, Knockdown, Western Blot, Expressing, Control, Phospho-proteomics, Quantitative RT-PCR

Journal: Frontiers in Microbiology

Article Title: Cyclooxygenase-2 Inhibition Reduces Autophagy of Macrophages Enhancing Extraintestinal Pathogenic Escherichia coli Infection

doi: 10.3389/fmicb.2020.00708

Figure Lengend Snippet: COX-2 up-regulation in ExPEC XM O2:K1:H7-infected macrophages through TLR4 mediated p38 and ERK1/2 MAPK pathway. (A) The TLR4 mRNA level of macrophages with or without ExPEC XM O2:K1:H7 treatment at 1 h was assayed by qRT-PCR. (B) Western blot analysis of COX-2 expression levels from cell lysates of RAW264.7 macrophages treated with or without TLR4 inhibitor (TAK-242) accompanied by ExPEC XM O2:K1:H7 infection for 6 h. (C) Western blot analysis of COX-2 expression levels from cell lysates of RAW264.7 macrophages treated with or without p38 MAPK inhibitor [SB202190 (FHPI)] and ERK1/2 MAPK inhibitor (SCH772984), accompanied by ExPEC XM O2:K1:H7 infection for 6 h. (D) Western blot analysis of indicated proteins with specific antibodies from cell lysates of RAW264.7 macrophages treated with or without 10 μM TAK-242, accompanied by ExPEC infection for 1 h.

Article Snippet: COX-2 inhibitor NS398 (50 μM), p38 MAPK inhibitor SB202190 (FHPI) (10 μM), and ERK1/2 MAPK inhibitor SCH772984 (10 μM), was purchased from Selleck Chemicals (Houston, United States).

Techniques: Infection, Quantitative RT-PCR, Western Blot, Expressing

Journal: bioRxiv

Article Title: ERK signaling licenses SKN-1A/NRF1 for proteasome production and proteasomal stress resistance

doi: 10.1101/2021.01.04.425272

Figure Lengend Snippet: A . Proteasome activity was reduced in SH-SY5Y Nrf1 shRNA cells compared to SH-SY5Y shRNA control cells, ****p<0.0001. B . SCH772984 treatment does not significantly reduce the background level of proteasome activity in HepG2 cells, ns, p>0.05. C . Screening of a commercial MAPK inhibitor library for small molecules that could increase the level of BTZ-induced apoptosis in HepG2 cells, as shown in D. Red and yellow indicate strong (robust CAS3 increase over control) and moderate (detectable CAS3 increase over control) hits, respectively. D . Induction of apoptosis in HepG2 cells that were treated with BTZ along with the indicated MAPK inhibitors from the library in C, with cell death detected by western blotting for cleavage of CAS3. E . Repeat analysis of “strong” hits from the screen shown in C and D.

Article Snippet: BTZ, SCH772984, and the MAPK inhibitor library were purchased from Selleck Chemicals.

Techniques: Activity Assay, shRNA, Control, Western Blot

Journal: Scientific Reports

Article Title: Marine alkaloid monanchoxymycalin C: a new specific activator of JNK1/2 kinase with anticancer properties

doi: 10.1038/s41598-020-69751-z

Figure Lengend Snippet: Analysis of the effect of ERK1/2 inhibitors FR180204 ( a ) and SCH772984 ( b ), MEK1/2 inhibitor PD98059 ( c ), or p38 inhibitor SB203580 ( d ) on MomC activity (Chou–Talalay method).

Article Snippet: The MEK inhibitor PD98059 was purchased from Merck Chemicals GmbH (Darmstadt, Germany), ERK1/2 inhibitors FR180204 and SCH772984 from Adooq bioscience (Irvine, CA, USA); PARP inhibitor olaparib JNK1/2 inhibitor SP600125 and p38 inhibitor SB203580 from LC Laboratories (Woburn, MA, USA).

Techniques: Activity Assay