Journal: Drug Design, Development and Therapy

Article Title: Inhibition of Pre-B Cell Colony Enhancing Factor Reduces Lung Injury in Rats Receiving Cardiopulmonary Bypass

doi: 10.2147/DDDT.S281554

Figure Lengend Snippet: Adenovirus-encoding sh-PBEF reduced the increased phosphorylation of ERK1/2, Akt, and p38MAPK in rat lung tissue. ( A ) Representative blots; ( B ) Phosphorylation of ERK1/2; ( C ) Phosphorylation of AKT; ( D ) Phosphorylation of p38MAPK vs control; Original blots were shown in Supplemental figure 1.*P<0.05; vs LPS, # P<0.05; vs CPB+Blank, @ P<0.05.

Article Snippet: The membrane was incubated with the following antibodies overnight at 4°C: rabbit anti-PBEF (1:2000, 11776-1-AP, Proteintech); rabbit anti-ERK1/2 (1:1000, 16443-1-AP, Proteintech); rabbit anti-p-ERK1/2 (1:500, bs-3016R, Bioss); rabbit anti-p38MAPK (1:1000, bs-0637R, Bioss); rabbit anti-p-p38MAPK (1:1000, bs-5477R, Bioss); rabbit anti-AKT (1:500, ab8805, Abcam); rabbit anti-p-AKT (1:1000, bs-2720R, Bioss); rabbit anti-AQP1 (1:1000, bs-1506R, Bioss); rabbit anti-AQP5 (1:1000, AF5169, Affinity); rabbit anti-ENaC (1:1000, bs-2957R, Bioss); rabbit anti-surfactant protein D (SP) (1:1000, ab220422, Abcam); and mouse monoclonal anti-GAPDH (1:2000, TA-08, ZSBio).

Techniques:

Journal: PLoS Genetics

Article Title: Regulation of Caenorhabditis elegans p53/CEP-1–Dependent Germ Cell Apoptosis by Ras/MAPK Signaling

doi: 10.1371/journal.pgen.1002238

Figure Lengend Snippet: (A) Phylogenic tree showing the evolutionary relationships of human (Hs), mouse (Mm), Drosophila (Dmel), C. elegans (Ce) and C. briggsae (Cb) MAPK phosphatases. LIP-1 is highlighted by the green oval within the DUSP6/7/9 cluster (blue oval). The other C. elegans MAPK phosphatase VHP-1 is highlighted by the grey oval. (B) In vivo dephosphorylation assays. Increasing amounts of Myc-tagged LIP-1 was expressed in Cos-1 cells and the phosphorylation of endogenous ERKs 1 and 2 (pERK), JNK (pJNK), or p38 (pp38) in response to either serum stimulation (ERK) or anisomycin (JNK and p38) was monitored by western blotting. Myc-tagged human DUSP5 and DUSP1 were used as positive controls to inactivate ERK and JNK/p38, respectively. (C) Sequence alignment of human DUSP6, DUSP7, DUSP9, and LIP-1, showing the conserved Kinase Interaction Motif (KIM). Increasing amounts of either Myc-tagged wild type or a mutant in which the conserved arginine residues (indicated by asterisks) of the KIM were mutated to alanine were expressed in Cos-1 cells and the phosphorylation of ERK1/2 was assessed. Tagged human DUSP1 was used as a positive control.

Article Snippet: Western blot analysis was performed using ERK (K-23, Santa Cruz 1∶2000, rabbit) and phosphorylated ERK (clone MAPK-YT, Sigma, 1∶2000, mouse) specific antibodies that cross react with MPK-1 and phosphorylated MPK-1 , , or anti-GLD-1 (1∶500, rabbit, this study) and HRP conjugated secondary antibodies (anti-rabbit-HRP and anti-mouse-HRP, DakoCytomation 1∶2000).

Techniques: In Vivo, De-Phosphorylation Assay, Western Blot, Sequencing, Mutagenesis, Positive Control

Journal: PLoS Genetics

Article Title: Regulation of Caenorhabditis elegans p53/CEP-1–Dependent Germ Cell Apoptosis by Ras/MAPK Signaling

doi: 10.1371/journal.pgen.1002238

Figure Lengend Snippet: (A and B) Wild type and let-60(ga89) and (C) wild type and let-60(n1046) worms were irradiated and allowed to recover at 20°C, and apoptotic corpses were scored after the stated time points. The scoring of apoptotic corpses was performed as in . (D) Activated (phosphorylated) MPK-1 levels differ between let-60(ga89) and let-60(n1046) worms. Protein was extracted from young adult worms (24 hours post L4 larval stage) and equal amounts were loaded onto SDS-PAGE gels. Activated MPK-1 was detected by an anti-P-ERK antibody, total MPK-1 by an anti-ERK antibody, and α-tubulin was used to control for loading. The ratio of phosphorylated MPK-1 to total MPK-1 (Ratio P∶T, normalised against wild type) is shown for the two isoforms.

Article Snippet: Western blot analysis was performed using ERK (K-23, Santa Cruz 1∶2000, rabbit) and phosphorylated ERK (clone MAPK-YT, Sigma, 1∶2000, mouse) specific antibodies that cross react with MPK-1 and phosphorylated MPK-1 , , or anti-GLD-1 (1∶500, rabbit, this study) and HRP conjugated secondary antibodies (anti-rabbit-HRP and anti-mouse-HRP, DakoCytomation 1∶2000).

Techniques: Irradiation, SDS Page

Journal: PLoS Genetics

Article Title: Regulation of Caenorhabditis elegans p53/CEP-1–Dependent Germ Cell Apoptosis by Ras/MAPK Signaling

doi: 10.1371/journal.pgen.1002238

Figure Lengend Snippet: (A and D) Wild type, (B and E) lip-1(gt448) , and (C and F) lip-1(zh15) worms (24 hours post L4 larval stage) were treated with 0 or 60 Gy of gamma irradiation and allowed to recover for two hours. Germlines were then dissected and processed for immunofluorescence using an anti-P-MPK-1 antibody (red). Nuclei were stained with DAPI (blue). All images show the germlines from the mid pachytene to the diakenesis region, the loop region (in which the cells are in late pachytene or early diplotene) is highlighted by the dotted arc, the mid pachytene region is highlighted by * and late pachytene by **. All germlines are orientated the same way: distal to the left and proximal to the right.

Article Snippet: Western blot analysis was performed using ERK (K-23, Santa Cruz 1∶2000, rabbit) and phosphorylated ERK (clone MAPK-YT, Sigma, 1∶2000, mouse) specific antibodies that cross react with MPK-1 and phosphorylated MPK-1 , , or anti-GLD-1 (1∶500, rabbit, this study) and HRP conjugated secondary antibodies (anti-rabbit-HRP and anti-mouse-HRP, DakoCytomation 1∶2000).

Techniques: Irradiation, Immunofluorescence, Staining

Journal: NPJ Precision Oncology

Article Title: Immunophenotyping with (phospho)protein profiling and fluorescent cell barcoding for single-cell signaling analysis and biomarker discovery

doi: 10.1038/s41698-024-00604-y

Figure Lengend Snippet: Antibody panel for (phospho)protein profiling

Article Snippet: p44/42 MAPK (pT202/Y204) , Cell signaling , 4375 , 0.5 μL + 9.5 μL.

Techniques: Control