MEASUREMENT

Overview

In order to evaluate the effect of our designed constructs on cell resilience, we performed a comparative survival assay involving Sf9 insect cells infected with different recombinant baculoviruses. The experiment measured post-freeze-drying recovery rates and quantified cell viability following rehydration.

Experimental Design

We conducted western blotting as our first step of measurement. Then， in phenotypic test,Fall armyworm cells infected with baculoviruses were divided into five experimental groups. The first group was infected with a baculovirus carrying an MCS insert, the second with SM, and subsequent groups with different constructs, with the final group serving as an uninfected control.

Prior to processing, cell viability was assessed using the trypan blue exclusion assay to count live versus dead cells. The cultures were then freeze-dried and stored under −80 °C in freezing medium.

Measurement and Analysis

After 24 hours of storage, samples were rehydrated, and cell viability was reassessed using trypan blue. The comparison between pre- and post-lyophilization cell counts allowed us to evaluate the survival rate of infected cells following freeze-drying and recovery.

Interpretation

By comparing cell survival among different infection conditions, we were able to assess the protective effects of MAHS, CAHS, and SAHS proteins during the freeze-drying process. This quantitative approach provided a clear measure of how tardigrade-derived proteins enhance cell resilience under extreme dehydration and rehydration cycles.