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Three other pathways which were altered to a lesser extent included: ERK/MAPK signalling, CDK5 signalling, and PI3K/AKT signalling (p = 0.016, 0.025, 0.039, respectively).
Using the less stringent dataset, five pathways were detected to be affected, among which the insulin receptor signalling (p = 0.047) (see Figure 3A).
The four pathways pertaining to cell signalling were signal transduction (p = 0.0009), cell-cell signalling (p = 0.012), protein amino acid phosphorylation (p = 0.015) and intracellular signalling cascade (p = 0.050).
Enriched canonical pathways included "leukocyte extravasation signalling" (p = 2.96E-05), "antigen presentation" (p = 3.9E-05 3.9E-05lper cell differentiation" (p = 1.74E-04), "dendritic cell maturation" (p = 5.96E-04) and "IL8 signalling" (p = 3.1E-03) (Table 4).
Endothelial cell contacts by junctional mechanisms (p =2.5e-2), regulatiof of translation initiation (p =2.6e-2), grAnzyme A signalling (p =2.9e-2), granzyme B signalling (p =3.1e-2), etc. emerged as significant processes.
Mechanistically, OLFM1 significantly downregulated p38 MAPK and caspase-3-dependent apoptotic signalling (p < 0.05; ESM Fig. 7).
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Among others, these included the following: Transport (p-value 3.42E-16), phosphorylationrylation (p-value 3.49E-13), Small GTPase-mediated signalling (p-value 1.94E-12) anDNA/RNANA processing (in particular, regulation of transcription, DNA-dependent, p-value 3.51E-08).
The anti P-STAT3 Tyr705 (Cell signalling), P-STAT3 Ser 727 (gift from Dr. Frank, [ 80], anti-STAT3 (F-2, Santa Cruz), anti-OCT4 (Abcam) and anti-ERK2 (Santa Cruz) antibodies were used as recommended by the manufacturer.
Among the 52 genes, 16 are involved in cell adhesion (p-value 7.4 × 10-11), and 14 are involved cell-cell signalling (p-value 7.9 × 10-6), suggesting a role in tumor invasiveness of the module [ 11].
The antibodies used were, SV40 TAg and IκBα from Santa Cruz Biotechnology, p53 and TnT from Calbiochem, phospho-AKT (T308), AKT, p-p70S6K (Ser389), p70S6K, p-AMPK (T172), AMPK, p-JNK-1/2 (T183/Y185), Cleaved Caspase 3, p-p42/44 (S202/T204) and p42/44 from Cell Signalling, p-IRS-1 from Millipore and p-Tyr, PAI-1, α- and β-actin and α-tubulin from Sigma Corp.
DETA-NONOate also activated ERK 1/2 signaling (p < 0.05).
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