In the present study, the authors performed a preliminary study to develop small molecule inhibitors of Ets transcription factors that play a crucial role in cancer progression. Their selection of target molecules appears appropriate. They have deployed two strategies, targeted immobilization NMR screening (TINS) and high throughput FRET screen, among which the latter gave more promising results: hit compounds from the HTS exhibited anti-angiogenic and anti-invasive activities in zebrafish models.
One of my concerns is that these two systems were independently evaluated. Comparison of the two strategies on the same platform would be informative for readers. I have a few suggestions to improve this manuscript as follows:

Major comments:
1) Compare the hit compounds (TINS and FRET) with some of previously developed Ets inhibitors reported in Ref 36-42 and discuss advantages and disadvantages of strategies deployed in this study. I noticed use of VPC-18005 in Figure 3D, but that is not enough.
2) How do the hit compounds in TINS behave in FRET screening system and vice versa?
3) Inhibitors obtained by the HTS are not specific to ETV6. How about those obtained from the TINS?
4) It would be interesting to see binding interfaces between EDBD and compounds obtained by the HTS. Is there a specific tendency?
5) Figure 3D: Include a cell line without aberration in Ets transcription factors as a negative control.

Minor comments:
6) In vitro DNA binding assay: Show sequences of oligonucleotide probes (Ets-binding and AP-1-binding).
7) Legend to Figure 1A: There are 6 lanes but there is no explanation. Are all these lanes necessary?
8) Figure 1C: A model depicting the EDBD-Fr 18 binding interface is slightly rotated and reduced in size. Should be corrected.
9) Figure 2A. middle: DMSO, 5 mM or 10 mM. Right?
10) In Fig. 2A, 2C, and 3A, letters are too small.
11) Legend to Figure 3D: Describe which cells have ERG or Fli1 fusion proteins. //

In this revised manuscript, the authors have addressed all my previous concerns. However, I have a minor comment on a newly added subpanel in Fig. 3D. In the subpanel, they examined the effect of YK-4-279 (an inhibitor of EWS-FLI1 fusion protein) on cell viability of VCaP cells containing an ERG fusion, but not SK-ES-1 cells containing a FLI1 fusion. I do not understand why they did not use SK-ES-1 cells. Some logical explanation would be helpful for readers.

In addition, line 469, “which is not unsurprising” may be “which is not surprising” or “which is unsurprising”.