Short hairpin RNA

effectively removing its concentration.

So if CD38, the expression of that protein on the surface, plasma membrane, is removed, deletion, right, double knockout, for example, or using a really powerful SI or SH RNA, and we find out that that protects against neurodegeneration or neuroinflammation, That means that with CD38 declining, there's more NAD.

See, that's the positive correlation.

But then I told you that NAD degradability isn't necessarily always linked simply to CD38.

R squared equaling 0.67, P being less than 0.0001.

The inhibition of tumor-originated SADM10 with its specific inhibitor GI254023X or its short hairpin RNA or SHRNA attenuated atrial fibrosis and AF susceptibility in breast cancer mice.

The authors conclude that breast cancer patients before treatment present a significantly higher prevalence of AF and atrial remodeling which correlates with elevated levels of tumour-originated SADM10 and ACF-secreted S-ephrin B2.

Tumour-originated SADM10 directly promotes AF pathogenesis by inducing atrial fibrosis independent of cancer treatment.

I guess something that may have been Maybe I was going to mention before in the methods, but that may be relevant for the results as well, is that we validated these findings orthogonally with multiple different approaches, especially the initial observation of the mistargeting of these axons and the loss of targeting the correct domain.

We not only did this pre-dependent loss of function at E15.5 in layer 2.3 neurons with the conditional knockout, but we also validated it with layer 2.3 specific delivery of shRNA of MEF2C and recapitulated the defect.

And additionally, after some suggestions were received, we validated this as well with double labeling of Cree positive and Cree negative axons in the same brain so that we were able to see within the same animal that the MEF2C knockout axons were cell autonomously unable to project to the right region.

but the wild type axons in the same brain that had some otherwise meant to see knockout axons were able to project to the correct region in the absence of any manipulations.

Perhaps the most surprising aspect of it was maybe just circling back to like double electroporations and just seeing that.

So in the initial electroporation, we tried to target like a broader scope of neurons.

So they were performed at around embryonic date 12.5. when layer 5 progenitors are migrating upwards of the cortex and those neurons that were being delivered the shRNA for the repulsive ligand FNA5 also had a cytosolic GFP tag and so we were able to see them with GFP and so just seeing specifically the rescue in the regions that had the GFP present and just seeing that the double manipulation specifically rescued this metastasic phenotype was very surprising Not so much surprising because the mechanism we had sort of worked out at the time, but it was really a nice thing to look at in New Mexico the first few times that we did.

Yeah, so in terms of surprises, that was definitely what's up there for me.