Extended Data Fig. 3: Gdf3 is not required for LPS-induced lethality in young mice but is regulated with age in a B cell-dependent manner, related to Fig. 1.

Figure S3 (A-E) Young (4- to 6-months-old) female WT and KO mice were injected with lethal dose of LPS (18 mg/kg) and VAT was collected for analysis 6 hr post-LPS injection. (A) Schematic of experimental design. (B) Frequency of ATM subsets in VAT. WT, n = 8; KO, n = 5. (C) Mean core BT after LPS injection. WT, n = 13; KO, n = 8. (D) Frequency of activated T cells and activated B cells in VAT. WT + PBS, n = 3; WT + LPS, n = 5; KO + LPS, n = 4. (E) Inflammatory gene expression in VAT. WT + PBS, n = 3; WT + LPS, n = 5; KO + LPS, n = 4. (F) Young (3-months-old) male WT and KO mice were injected with rGDF3 (10 µg/kg) 24 hours prior to lethal dose of LPS (18 mg/kg). Mean core BT was measured after LPS injection (left). VAT was collected for analysis 6 hr post-LPS injection (right). (G) TGFβ superfamily gene expression in VAT and (H) Gdf3 expression in multiple tissues from unchallenged 2-months-old (n = 4) and 21-month-old (n = 4) female WT mice. (I) TGFβ superfamily gene expression in VAT from 20- to 22-month-old female WT (n = 6) and B^null (n = 9) mice injected with 0.1 mg/kg LPS. (J-M) Metabolic profiling of old (21- to 26-month-old) control and mKO mice. Female control, n = 11; female mKO, n = 9; male control, n = 8; male mKO, n = 5. Body weights and glucose tolerance test of female (J-K) and male (L-M) mice. (N-R) Old (21- to 24-month-old) female control and mKO mice were injected with 0.1 mg/kg LPS. Frequency of (N) CD9⁺ ATMs, (O) CD206⁺ ATMs, and (P) CCR2⁺ ATMs in VAT. (Q) Frequency of CD11c⁺ and CD206⁺ ATMs in SAT. Control, n = 7 ~ 14; mKO, n = 4 ~ 6. (R) Free fatty acid released from VAT explants from old female control (n = 5) and mKO (n = 4) mice. (S) Frequency of CD11c⁺ ATMs from VAT of male and female mice injected with 0.1 mg/kg LPS. Females ♀, males ♂. Control, n = 14; mKO, n = 9. (T) Representative gating strategy for B cells. (U-V) Frequency of (U) CD73⁺IgM⁺ B cells and (V) PDL1⁺ B cells in VAT of old control and mKO mice injected with 0.1 mg/kg LPS. (W) Mean core BT of young (3-month-old) or old (23- to 26-month-old) male mice post-LPS (0.1 mg/kg) injection. Young control, n = 8; old control n = 8; old mKO, n = 5. All data are presented as means ± SEM.