Those with a great memory will remember that I’ve been struggling (1, 2, 3) to identify adherens and tight junction proteins (i.e., cadherins and ZO-1) in bEnd3 endothelial cells. Since I’ve recently moved over to investigate air-blood barrier co-culture models,…
Author: Barrett Nehilla
Air-blood barrier #1
Our collaboration with Alison Elder, et al. in the Dept. of Environmental Medicine has led us to talk about developing a co-culture model of the air-blood barrier. This barrier is ‘located’ at the alveolar sacs in the lungs, where lung…
Posted in NRG
Vacuoles via SEM
I had some extra samples from last week and had been wanting to try SEM on cells, so I went for it this week. Specifically, I was hoping to get images of vacuoles in cells over free-standing pnc-Si. I followed…
Posted in NRG
Amyloid-Beta separations
I’ve been working recently with amyloid-beta (AB) provided to me by some people in Brad Nilsson’s lab. These experiments are related to the HMM diffusion experiments for our collaboration with Harold Smith. Basically, AB fibrils are very long (microns) and…
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More vacuoles on free-standing than supported pnc-Si (and PET)
One of the other suggestions from Angela Glading at lab meeting a while ago was to quantify the vacuoles on different culture areas. So, I selected images from 3 different experiments (all at day14 of growth) and counted the number…
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HMM vs. LMM diffusion
My last post dealing with this project was the first to definitively demonstrate HMM diffusion through pnc-Si. Unfortunately, there wasn’t a difference between the HMM filtrate signal for intact and digested (+RNAse) samples. As a reminder, HMM is a large…
Epithelium on transwells(2)
My last post was the first experiment I performed with rat lung epithelial cells on transwells. It seemed like there was vacuolization but it was delayed compared to endothelial cells (and only happened on 1 of 2 pnc-Si transwells). Here,…
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Lung epithelial cells on transwells(1)
A couple weeks ago when I presented at lab meeting, Angela Glading suggested that I look at epithelial cell vacuolization on transwells. This was in order to determine if vacuole formation was endothelial cell-specific. If I’m remembering correctly, the rationale…
Posted in NRG
HMM Diffusion through pnc-Si Sepcons
Carrie and I have been trying to detect HMM (APOBEC3G) diffusion through Sepcons without much luck. We determined that the dilution factor (~1:30) for diffusion in our 384-well plate format was too high to detect HMM in the basolateral well. …
Posted in NRG
Diffusion through non-porous membranes figure
A couple weeks ago, I presented a potential figure for my paper that didn’t make sense. So, I did a couple more controls and put together this new figure. This is the “diffusion through ‘non-porous’ SC348/cut-off of nonporous membranes” figure. …
Posted in NRG
