Fig 1.
Summary of experiments included in this lab protocol.
Fig 2.
Rearing and maintenance of G. mellonella in the science research laboratory.
(a) Housing equipment for G. mellonella. (b) The lifecycle of G. mellonella reared and maintained in our research laboratory. The larval stage of G. mellonella (eggs to last-instar stage larvae) was maintained at a temperature-controlled condition with a heating mat.
Fig 3.
G. mellonella infection assays.
(a) Healthy larvae have a white-creamy appearance with no melanisation. Dead infected larvae showed different levels of melanisation and were unresponsive to physical stimuli. (b) Kaplan-Meier survival curves of the killing assay. The results showed that A. baumannii strain C98 killed G. mellonella dose-dependently. (c) Bar chart of bacterial growth in vivo. The results demonstrated that A. baumannii strain C98 increased rapidly in the G. mellonella after 3 hours of incubation.
Fig 4.
Bacterial RNA isolation from G. mellonella larvae infected by A. baumannii strain C98.
(a) Steps of bacterial cell harvesting from the infected larvae. The host cells were removed by multiple centrifugations to minimize the contaminations of the host’s RNA. (b) Quality assessment of RNA samples using Agilent 2200 TapeStation System and the RIN numbers were calculated by TapeStation Software A.02.02. A0: Electronic ladder; A1: bacterial RNA from the mid-exponential phase in broth culture (control); B1: bacterial RNA from infected larvae after 3-hour infection. (c) ImageJ quantification of bacterial RNA from the infected larvae (B1). Ratios of 23S:16S were calculated by the Area Under the Curve to assess the RNA integrity.