Mouse anti yap sc 101199

Proteins were extracted from neuroblastomas cells, and then separated by SDS-PAGE and transferred to a polyvinylidene difluoride membrane (PVDF). After blocked with 5% nonfat milk in TBST for 2 hours, the membrane was incubated with primary antibodies. Membranes were washed three times and incubated with the horseradish peroxidase-conjugated second antibodies. The signals were captured by the ECL reagent (Beyotime) and visualized by western blotting detection instruments (Clinx Science). Mouse anti-TAZ (560235; BD Biosciences), mouse anti-YAP (sc-101199, 1:200), goat anti-CTGF (sc-14939, 1:200) from Santa Cruz company, rabbit anti-PDGF-β (E1A0240-1, 1:1000) from EnoGene, and mouse anti-GAPDH (AG019, 1:1000) from Beyotime, cell cycle regulation antibody sampler kit #9932 from Cell Signaling Technology were used as primary antibodies. HRP-labeled goat anti-mouse IgG (H + L) (A0216, 1:5000) and goat anti-rabbit IgG (H + L) (A0208) were used as secondary antibodies which purchased from Beyotime.

Mouse anti-vinculin (V9131, dilution ratio: 1/20,000 (WB), 1/500 (IF)), rabbit anti-CAP (SORBS1) (HPA027559, 1/2,000 (WB), 1/100 (IF)) and anti-β tubulin (T4026, 1/2,000) antibodies were purchased from Sigma (Saint Louis, MO). Mouse anti-β-actin antibody (ab6276, 1/10,000) was purchased from Abcam (Cambridge, UK). Mouse anti-YAP (sc-101199, 1/100 (IF), 1/1000 (WB)), and rabbit anti-ERK2 (sc-154, 1/10,000) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). Rabbit anti-aP2 (#3544, 1/2,000) antibodies was purchased from Cell Signaling Technology (Boston, MA). Rabbit anti-vinexin and anti-ArgBP2 polyclonal antibodies were described previously^{18 (link),22 (link)}. Alexa Fluor 568 phalloidin and 633 phalloidin was purchased from Thermo Fisher Scientific (Rockford, IL). Type I collagen was purchased from Nitta Gelatin (Osaka, Japan). Insulin and 3-Isobutyl-1-methylxanthine (IBMX) were purchased from Sigma.

All cell lines were purchased from the ATCC and cultured under standard conditions. All cell-based assays were performed as described (Nguyen et al. 2017 (link); Nguyen et al. 2016 (link)). YAP localization was scored using mouse-anti-YAP (sc-101199, Santa Cruz) as previously described (Nguyen et al. 2017 (link)). Antibodies to phospho-YAP Ser127 (Cat #4911), YAP (#4912), TAZ (#2149), LATS2 #5888), phospho-LATS1/2 (#8654), Myc-Tag (#2272), LKB1 (#3050), MARK1 (#3319) and p-MARK1-4 (#4386) were from Cell Signaling. YAP (#sc-101199) antibody used for immunohistochemical staining was obtained from Santa Cruz Biotechnology. Anti-FLAG (M2, #F3165) and RhoBTB2 (SAB1407189) were from Sigma. Anti-actin (#MAB1501) was from Millipore.