Extended Data Fig. 6: Basolateral infection of human organoids by MRV generates a stronger intrinsic immune response compared to apical infection.
(a) Schematic of the microinjection approach used to infect organoids specifically from either their apical or basolateral sides. (b-c) Human intestinal organoids were infected by MRV apically or basolaterally through microinjection. Virus infection was assessed 16 hpi for (b) the production of the non-structural viral protein μNS using Western blot analysis. Representative western blot is shown. and (c) the production of the MRV μ2 genome segment by q-RT-PCR. Results shown the mean +/- sd. n=3. (d) Generation of intrinsic immune response was assessed by quantifying the production of type I (IFNβ) and type III (IFNλ2/3) IFNs using q-RT-PCR. Results shown the mean +/- sd. n=3. (c-d) P values were calculated with a two-tailed unpaired t test using GraphPad Prizm. (e) The relative expression of TLR2-9 was evaluated in human intestinal organoids. Results are expressed as a ratio to the house keeping gene HPRT1. Results shown the mean +/- sd. n=3.
