MONJU is equipped with hydrogen gas detectors for the small water leak detection, gas pressure gauges for the medium leak and sensors of rupture discs for the large leak.
Although as the reviewers have commented that "…the cell might create a hole in a substrat…," we speculate there are two explanations for the production of these "holes" (area lack of fluorescent signal, see Author response image 1A): (1) NP-conjugated DNA strain on 56 pN NP-TGT sensors were ruptured and dissociated from the coverslip.
These results suggest that the internalization of BCR and antigen molecules did not contribute to the different levels of the accumulation of the IgM-BCRs into the B cell IS in response to different NP-TGT sensors with different mean rupture forces.
In this report, we modified the dsDNA-based TGT developed by Ha and his colleagues (Wang and Ha, 2013) to acquire a series of 8 NP-TGT sensors with predefined mean rupture force from low to high, respectively (12, 16, 23, 33, 43, 50, 54 and 56 pN at the condition in the original study).
In our assessment, this is due to a rupture of the various sensor films from the effects of shear force.
For the integrin liposomes, we obtained an almost linear relationship after the equilibration period, which indicates that the liposomes did not rupture on the SiO2 sensors.
Supported by NP-TGT system, for the first time, we demonstrated that IgM-BCR activation is in a mechanical force dependent manner by providing evidence that high mean rupture force NP-TGT sensors which can stand for high-force load-triggered more aggressive spreading response and the enhanced BCR signaling than low mean rupture force NP-TGT sensors.
We also agree with the reviewer that the best guess of the relevant time scale of rupturing single NP-TGT sensor in our experimental system would be a seconds to dozens of seconds.
In addition, we would like to share with the reviewers our thoughts that direct comparison between the rupture force of TGT sensors (Wang X and Ha T, Science, 2013, 340 991-4) and the rupture force between NP hapten antigen and B1-8 BCR (Natkanski et al., Science, 2013, 340 1587-90) should be avoided.
Then what should the time scale of rupturing a single NP-TGT sensor at the physiological condition be?
