palk y1604 #3341 antibody  (Cell Signaling Technology Inc)

Journal: Cancers

Article Title: A Polytherapy Strategy Using Vincristine and ALK Inhibitors to Sensitise EML4-ALK-Positive NSCLC

doi: 10.3390/cancers14030779

Figure Lengend Snippet: The combination of ALK inhibitor and vincristine impairs downstream signalling of RAS/MAPK, PI3K/AKT and JAK/STAT3 in H3122 but not H2228 cells. ( A ). H3122 and H2228 cells were serum starved overnight and then treated with the indicated inhibitors for 3 h (crizotinib/ceritinib 400 nM and vincristine 20 nM). Western blotting analysis for the indicated proteins was performed. β-actin was used as a loading control. ( B – E ). ELISA assay was used to quantify expressions of pALK Y1604 , pSTAT3 Y705 , pAKT S473 and pERK T202/Y204 proteins. Data represent counts from three biological replicates, n = 3. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 in comparison to DMSO by one-way ANOVA. ns = not significant.

Article Snippet: Immunoblot analysis were performed with the following primary antibodies: pALK (Y1604) (1:1000, 3342; Cell Signaling Technology), ALK (1:1000, 3791; Cell Signaling Technology), pSTAT3 (Y705) (1:1000, 9145; Cell Signaling Technology), STAT3 (1:1000, 9139; Cell Signaling Technology), pAKT (S473) (1:1000, 9271; Cell Signaling Technology), AKT (1:1000, 9272; Cell Signaling Technology), pERK (T202/Y204) (1:1000, 9101; Cell Signaling Technology), ERK (1:1000, 9102; Cell Signaling Technology), Acetyl-α-tubulin (Lys40) (1:1000, T7451; Sigma-Aldrich), GAPDH (1:2000, ab37168; abcam) and β-actin (1:10,000, 5441; Sigma-Aldrich, St. Louis, MO, USA) ( ).

Techniques: Western Blot, Control, Enzyme-linked Immunosorbent Assay, Comparison