Figure 3.
Eat-me signals and receptors involved in infection-induced non-professional efferocytosis. (A and B) Heat map of the fold increased expression of selected eat-me receptor, mediator, and signal genes in total isolated IECs from healthy neonatal (day 3 after birth) versus healthy adult (day 21 after birth) mice (A) and neonatal 1-day-old mice infected with 102 CFU S. Typhimurium (S. Tm) at day 4 p.i. versus healthy age-matched uninfected neonatal mice (B). n = 4. (C) Heat map of the fold increased expression of selected eat-me receptor, mediator, and signal genes in flow cytometrically sorted Salmonella-positive (Salmonella+) and Salmonella-negative (Salmonella−) EpCam+ IECs obtained from infected neonatal mice versus total isolated IECs obtained from age-matched uninfected mice. 1-day-old mice were infected with 102 CFU constitutively mCherry expressing S. Typhimurium. Total small IECs were isolated at day 4 p.i. and sorted by flow cytometry to obtain EpCam+ CD45−Salmonella-positive (mCherry+) and EpCam+ CD45−Salmonella-negative (mCherry−) cells. Total RNA of the described cell populations was obtained from a total of 49 neonatal animals from 7 litters. RNA samples from the same population were pooled into one sample for analysis, n = 1. (D–H) Identification of eat-me receptors, mediators, and signals involved in non-professional efferocytosis. Neonatal stem cell organoid cell monolayers were preincubated with an anti-CD36 antibody or an isotype control antibody (D), an anti-integrin-αv (CD51) antibody or an isotype control antibody (E), or an anti-EpCAM antibody (H) for 60 min. Alternatively, stem cell organoid–derived starved IECs were preincubated with C1q (F) or annexin V (G) for 15 min. After 2 h coculture of pretreated cell monolayers with untreated starved IECs (D, E, and H) or untreated cell monolayers with pretreated starved IECs (F and G), efferocytosis events (TR+ cells) were recorded in 20–40 image fields of the size 1.25 × 1.00 mm obtained in two independent experiments. Mann–Whitney U test. ***, P < 0.001; ****, P < 0.0001; ns, non-significant.

Eat-me signals and receptors involved in infection-induced non-professional efferocytosis. (A and B) Heat map of the fold increased expression of selected eat-me receptor, mediator, and signal genes in total isolated IECs from healthy neonatal (day 3 after birth) versus healthy adult (day 21 after birth) mice (A) and neonatal 1-day-old mice infected with 102 CFU S. Typhimurium (S. Tm) at day 4 p.i. versus healthy age-matched uninfected neonatal mice (B). n = 4. (C) Heat map of the fold increased expression of selected eat-me receptor, mediator, and signal genes in flow cytometrically sorted Salmonella-positive (Salmonella+) and Salmonella-negative (Salmonella−) EpCam+ IECs obtained from infected neonatal mice versus total isolated IECs obtained from age-matched uninfected mice. 1-day-old mice were infected with 102 CFU constitutively mCherry expressing S. Typhimurium. Total small IECs were isolated at day 4 p.i. and sorted by flow cytometry to obtain EpCam+ CD45−Salmonella-positive (mCherry+) and EpCam+ CD45−Salmonella-negative (mCherry−) cells. Total RNA of the described cell populations was obtained from a total of 49 neonatal animals from 7 litters. RNA samples from the same population were pooled into one sample for analysis, n = 1. (D–H) Identification of eat-me receptors, mediators, and signals involved in non-professional efferocytosis. Neonatal stem cell organoid cell monolayers were preincubated with an anti-CD36 antibody or an isotype control antibody (D), an anti-integrin-αv (CD51) antibody or an isotype control antibody (E), or an anti-EpCAM antibody (H) for 60 min. Alternatively, stem cell organoid–derived starved IECs were preincubated with C1q (F) or annexin V (G) for 15 min. After 2 h coculture of pretreated cell monolayers with untreated starved IECs (D, E, and H) or untreated cell monolayers with pretreated starved IECs (F and G), efferocytosis events (TR+ cells) were recorded in 20–40 image fields of the size 1.25 × 1.00 mm obtained in two independent experiments. Mann–Whitney U test. ***, P < 0.001; ****, P < 0.0001; ns, non-significant.

This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal