Journal: Nature Immunology
Article Title: Dynamic mitochondrial transcription and translation in B cells control germinal center entry and lymphomagenesis
doi: 10.1038/s41590-023-01484-3
Figure Lengend Snippet: a , Schematic depicting the adoptive transfer strategy of lymphoma cells from Eμ-Myc mice. After the development of lymphoma, cells were transferred into WT congenic (CD45.1 + ) recipients. Representative flow cytometry plots showing the identification of CD19 + CD45.2 + donor lymphoma cells and CD19 + CD45.1 + WT B cells from the inguinal lymph nodes of recipient mice after 3 weeks. b , Flow cytometry histogram plot depicting COXI expression in CD45.2 + lymphoma cells and CD45.2 − WT B cells ( n = 5 mice) as described in a . FMO, fluorescence minus one. Quantification of COXI gMFI. Data are representative of two independent experiments. c , Flow cytometry histogram depicting TFAM expression in CD45.2 + lymphoma cells and CD45.1 + WT B cells ( n = 5 mice) as described in a . Quantification of TFAM gMFI. Data are representative of two independent experiments. d , Kaplan–Meier graph depicting the survival curve of Eμ-Myc × Tfam homozygous ( Cd79a Cre/+ Tfam loxP / loxP × Eμ-Myc, n = 9), Eμ-Myc Tfam heterozygous ( Cd79a Cre/+ Tfam loxP /+ × Eμ-Myc, n = 13) and Eμ-Myc ( Cd79a +/+ Tfam loxP / loxP × Eμ-Myc or Cd79a +/+ Tfam loxP /+ × Eμ-Myc, n = 18) mice followed for up to 6 months of age. e , Representative flow cytometry histogram depicting COXI expression in the Daudi cell line treated with the IMT1 at various concentrations (0.1 μM, 1 μM, 10 μM) or vehicle (DMSO) for 120 h. Representative of three independent experiments. f , Quantification of COXI levels by flow cytometry, normalized to SDHB in IMT1- or vehicle-treated Daudi cells at different concentrations and time points (0, 24, 48, 72, 96, 120 h), representative of two independent experiments. g , Airyscan ICC confocal images of COXI and TOMM20 in Daudi cells treated with DMSO or IMT1 (1 μM) for 120 h. Scale bar, 3 μm. Representative of two independent experiments. h , Quantification of live Daudi cell concentrations over a 120-h incubation period in the presence of IMT1 or vehicle (DMSO), representative of two independent experiments with three to five technical replicates each. i , Flow cytometry-based quantification of mtROS in the Daudi cell line treated with IMT1 (1 μM) or vehicle (DMSO) for 120 h. Each point represents technical replicates ( n = 3 per group). Representative of two independent experiments. j , Airyscan confocal images of Daudi cells treated with IMT1 (1 μM) or DMSO and stained for F-actin phalloidin. Scale bar, 5 μm. Representative of two independent experiments. k , Quantification of F-actin phalloidin fluorescence by flow cytometry ( n = 5 technical replicates). Data are representative of two independent experiments. l , Quantification of COXI levels in Daudi cells by flow cytometry after treatment with vehicle (ethanol) or CHL at 10 μg ml −1 or 25 μg ml −1 for 120 h ( n = 8 technical replicates). Data are representative of two independent experiments. m , Quantification of F-actin levels in Daudi cells by flow cytometry after treatment with vehicle (ethanol) or CHL at 10 μg ml −1 or 25 μg ml −1 for 120 h ( n = 8 technical replicates). Data are representative of two independent experiments. n , Quantification of live Daudi cell concentrations over a 120-h incubation period in the presence of CHL at 10 μg ml −1 or 25 μg ml −1 or vehicle (ethanol) as in m . Representative of two independent experiments with four technical replicates ( n = 4). Statistical significance was calculated using an unpaired two-tailed t -test ( b , c , i ), a Mantel–Cox log-rank test ( d ) or an ordinary one-way ANOVA with Tukey’s ( h , n ) or Dunnett’s multiple comparisons test ( k – m ). Data are presented as the mean ± s.e.m.
Article Snippet: B6.Cg- Tfam tm1.1Ncdl /J (strain 026123), B6.C(Cg)- Cd79a tm1(Cre)Reth /EhobJ (strain 020505), B6.129P2- Aicda tm1(Cre)Mnz /J (strain 007770), B6;129S6- Gt(ROSA)26Sor tm9(CAG- tdTomato)Hze /J (strain 007905), B6.Cg-Tg(TcraTcrb)425Cbn/J (strain 004194) and B6.Cg-Tg(IghMyc)22Bri/J (strain 002728) were purchased from The Jackson Laboratory.
Techniques: Adoptive Transfer Assay, Flow Cytometry, Expressing, Fluorescence, Incubation, Staining, Two Tailed Test
