Fig. 6: Blocking Tim4 impairs post-prandial cholesterol metabolism.

a Mice were kept on CD or fed with HFD overnight. Mice fed HFD were injected i.p. with Ig control or blocking anti-Tim4 Ig or chloroquine, 72 h and on the day prior to the overnight HFD. b Flow cytometric analysis on ATMs showing F4/80^highTim4⁺, F4/80^highTim4⁻, and F4/80^low ATM populations and quantification of their relative proportions in mice kept on CD (gray bar) and mice fed overnight with HFD and injected with Ig control (red), anti-Tim4 Ig (blue), or chloroquine (green). Data are representative of 2 independent experiments with n = 5 mice per groups. c–e Circulating levels of NEFA (c), total cholesterol, HDLc, and non-HDLc (d), and TG (e) in mice kept on CD (gray bar) and mice fed overnight with HFD and injected with Ig control (red), anti-Tim4 Ig (blue), or chloroquine (green). f–h, j Flow cytometric analysis showing histogram of lysotracker (f), CD36 (g), ABCA1 (h), and HSP70 (j) fluorescence intensity in F4/80^highTim4⁺ ATMs and quantification of MFI in the indicated ATM populations from mice kept on CD (gray bar) and mice fed overnight with HFD and injected with Ig control (red), anti-Tim4 Ig (blue), or chloroquine (green). To allow detection of Tim4 in mice receiving anti-Tim4 Ig, Tim4 was detected using rat anti-Tim4 Ig plus secondary anti-Rat Ig-647. Data are pooled from n = 8–15 mice per group from 2 to 3 independent experiments. i Ratio of relative amounts of Abca1 and Nr1h3 expressed by F4/80^highTim4⁺ ATMs isolated from mice kept on CD or fed with HFD overnight and treated with Ig control or blocking anti-Tim4 Ig as in (a). Data are pooled from n = 3 biological replicates per group. Error bars show SEM. Kruskal–Wallis test with Dunn’s multiple comparisons test or ANOVA with Sidak’s multiple comparisons test were applied after assessing normality using D’Agostino and Pearson Normality test (b–h, j). Two-tailed Student’s T test was applied in (i). Significant differences are indicated by *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, ns = non-significant.