Skip to main content
. 2019 Apr 12;20(6):e47227. doi: 10.15252/embr.201847227

Figure EV2. cTAZ was not regulated by Hippo signaling.

Figure EV2

  1. Protein domains and interacting proteins of TAZ. Phosphorylation sites of LATS kinases are shown.
  2. cTAZ did not interact with LATS2. HEK293A cells were transfected with Flag‐tagged cTAZ, TAZ, YAP, WW‐mutated TAZ (WW), and WW1/2‐mutated YAP (WW1/2), and cell lysates were prepared and used for IP.
  3. cTAZ was not regulated by serum stimulation. cTAZ‐ or TAZ‐overexpressing MCF‐10A cells were stimulated by 10% FBS for 1 h, and protein expression was determined. Ectopic and endogenous TAZ were down‐shifted upon serum stimulation, whereas no change for cTAZ was detected.
  4. cTAZ was not regulated by serum starvation. cTAZ‐ or TAZ‐overexpressing HEK293A cells were serum‐starved for 8 h and harvested for IB. The phosphorylation of full‐length TAZ was also determined using phos‐tag gel. The asterisk indicates a shift of TAZ on regular gel.
  5. Deletion of LATS kinases stabilized TAZ, but not cTAZ. WT or LATS1/2‐double‐knockout (dKO) HEK293A cells were transfected with Flag‐tagged cTAZ or TAZ. Cells were treated with CHX (50 μg/ml) in a time course (0–4 h), and protein levels were determined. Error bars indicate SD, n = 3. ***P < 0.001; two‐way ANOVA test was used for statistical analysis.
  6. Serum treatment stabilized TAZ, but not cTAZ. RKO cells cultured in the presence or absence of 10% serum were treated with CHX (50 μg/ml) in a time course (0–6 h), and protein levels were determined. Error bars indicate SD, n = 3. ***P < 0.001; two‐way ANOVA test was used for statistical analysis.

Source data are available online for this figure.