Journal: Cell Reports Medicine

Article Title: Therapeutic activity of a hematopoietic stem cell-delivered cell-penetrating frataxin in Friedreich’s ataxia models

doi: 10.1016/j.xcrm.2026.102803

Figure Lengend Snippet: LV-FXN gene therapy does not affect the engraftment and lineage commitment of HSPCs and deposits FXN protein in FRDA-relevant tissues (A) Overview of the transplantation experiment. Lineage-negative cells isolated from LY5.1 mice were transduced with LV-FXN at a multiplicity of infection (MOI) of 20 and transplanted into lethally irradiated LY5.2 recipient mice. Three months after transplantation, hematopoietic organs were analyzed by FACS, and frataxin protein levels were measured in the spleen, brain, heart, muscle, liver, and kidney by mass spectrometry. (B–D) Percentage of CD45.1 (donor-derived) and CD45.2 (recipient-derived) cells in peripheral blood, bone marrow (BM), and spleen of mice transplanted with mock-untransduced ( n = 3) or LV-FXN-transduced cells ( n = 4) (upper), along with the lineage composition within the CD45.1 and CD45.2 compartments (lower). (E) Levels of human mature frataxin (ng per mg of total protein; mean ± SD) in the spleen of mice transplanted with mock-untransduced cells (mouse #304) or LV-FXN-transduced cells (mice #306, #307, and #310). (F) Total frataxin levels (ng per mg of total protein; mean ± SD) in the indicated organs of mice transplanted with mock-transduced cells (mouse #304) or LV-FXN- transduced cells with >1 vector copy number (VCN) (mice #307 and #310). ND = not determined.

Article Snippet: Anti-mouse CD45.1 PE , Miltenyi , Cat# 130-102-499; RRID:AB_2660704.

Techniques: Transplantation Assay, Isolation, Transduction, Infection, Irradiation, Mass Spectrometry, Derivative Assay, Plasmid Preparation

Journal: iScience

Article Title: A distinct TCR + CD19 + population in the peritoneal cavity: B1a cells as precursors of extrathymic T cells

doi: 10.1016/j.isci.2026.115505

Figure Lengend Snippet: Adoptively transferred B1a cells differentiate into DP cells and extrathymic T cell lineages (A) Two weeks after the adoptive transfer of B1a cells into muMT mice, the frequencies of TCR + CD19 + DP cells and B1a cells were assessed in the PEC, spleen, liver, small intestine, and thymus by flow cytometry. (B) Four weeks after the adoptive transfer of CD45.2 + B1a cells into CD45.1 + recipients, flow cytometric analysis was performed to evaluate the frequencies of CD45.2 + extrathymic T cells, DP cells, and B1a cells in the indicated tissues.

Article Snippet: CD45.1 + CD3 + T cells were isolated from mouse spleens, and CD45.2 + CD19 + B cells were isolated from the peritoneal cavity using magnetic-activated cell sorting (MACS) with anti-biotin microbeads (130-097-046; Miltenyi Biotec) according to the manufacturer’s instructions.

Techniques: Adoptive Transfer Assay, Flow Cytometry

Journal: bioRxiv

Article Title: Stem cell function in vivo is supported by an alternative glycolysis endpoint

doi: 10.64898/2026.03.30.715412

Figure Lengend Snippet: A, Schematic of canonical glycolysis terminating via fermentation of glucose to lactate by lactate dehydrogenase (LDH) or via conversion of pyruvate to acetyl-CoA by pyruvate dehydrogenase (PDH) for oxidation in the tricarboxylic acid (TCA) cycle. B, Immunoblot showing loss of LDHA and LDHB proteins in bone marrow cells following ACK lysis to remove red blood cells, 3 weeks after Mx1Cre induction with poly(I:C). C–E, The number of bone marrow HSCs in two legs in Mx1Cre;Ldha Δ/Δ (c), Mx1Cre;Ldhb Δ/Δ ( D ), and Mx1Cre;Ldha Δ/Δ ;Ldhb Δ/Δ ( E ) mice and littermate controls at 3 and 6 weeks after deletion. F–G, Competitive BM transplantation of donor CD45.2 + Mx1Cre;Ldha Δ/Δ , Mx1Cre;Ldhb Δ/Δ , or Ldha fl/fl or Ldhb fl/fl littermate control bone marrow cells with wild-type CD45.1;CD45.2 competitor cells to lethally irradiated CD45.1 recipient mice. Shown are donor chimerism in F peripheral blood and G BM HSPCs (n = 12–15 mice per genotype for blood; n = 10–14 for BM; three independent experiments). H–I, Competitive transplantation of Mx1Cre;Ldha Δ/Δ ;Ldhb Δ/Δ or littermate control BM cells. Donor contribution to H peripheral blood over 16 weeks and I BM HSPCs at week 16 (n = 11–15 for blood; n = 10–14 for BM; three independent experiments). J–K, Secondary transplantation of 5 × 10⁶ BM cells from primary recipients shown in H-I . Donor chimerism in J peripheral blood and K BM HSPCs (n = 13–14 from three independent experiments). L–M, Competitive transplantation of undeleted donor Mx1Cre;Ldha fl/fl ;Ldhb fl/fl or Ldha fl/fl ; Ldhb fl/fl littermate control bone marrow cells followed by deletion at 6 weeks. Donor chimerism in l blood and M BM HSPCs (n = 11–15 from three independent experiments). Data represent mean ± s.d. Statistical significance was assessed with a 1-way ANOVA ( F-G ) followed by Dunnett’s test, or with a t-test (rest). All figures show * p<0.05, ** p<0.01, ***p<0.001.

Article Snippet: For competitive transplantation experiments, 500,000-1,000,000 cells each of donor (CD45.2) and competitor (CD45.1/CD45.2) cells were mixed and transplanted into the retro-orbital venous sinus of anesthetized CD45.1 recipients irradiated using an XRAD 320 X-ray irradiator (Precision X-Ray) with two doses of 540 rad (1080 rad in total) at least 3 hours apart.

Techniques: Western Blot, Lysis, Transplantation Assay, Control, Irradiation

Journal: bioRxiv

Article Title: Stem cell function in vivo is supported by an alternative glycolysis endpoint

doi: 10.64898/2026.03.30.715412

Figure Lengend Snippet: Transplantation of Mx1Cre;Ldha Δ/Δ ; Ldhb Δ/Δ ;Pdha1 τι , Mx1Cre;Ldha Δ/Δ ;Pdha1 τι , or littermate control CD45.2 + donor bone marrow cells with wild type CD45.1 + CD45.2 + competitor cells into lethally irradiated CD45.1 + recipients. A, Schematic of primary and secondary transplantation experiments. B, Schematic of transplantation of undeleted donor cells followed by deletion at 6 weeks post-transplant. C–D, Donor chimerism in C blood and D bone marrow HSPCs in primary recipients (n = 12-13 for blood; n = 9-12 for BM from 3 independent experiments) from the transplant outlined in A . E–F, Donor contribution in E blood and F bone marrow HSPCs in secondary recipients (n = 8-11 for blood; n = 4-8 for BM from 3 independent experiments) from the transplant outlined in A . G–H, Peripheral blood ( G ) and BM HSPC ( H ) chimerism (n = 20–29 for blood; n = 13–24 for BM from 6 independent experiments) from the transplant outlined in B. All data represent mean ± s.d. Statistical significance was assessed with a 1-way ANOVA followed by Dunnett’s test. All figures show * p<0.05, ** p<0.01, ***p<0.001.

Article Snippet: For competitive transplantation experiments, 500,000-1,000,000 cells each of donor (CD45.2) and competitor (CD45.1/CD45.2) cells were mixed and transplanted into the retro-orbital venous sinus of anesthetized CD45.1 recipients irradiated using an XRAD 320 X-ray irradiator (Precision X-Ray) with two doses of 540 rad (1080 rad in total) at least 3 hours apart.

Techniques: Transplantation Assay, Control, Irradiation

Journal: bioRxiv

Article Title: Stem cell function in vivo is supported by an alternative glycolysis endpoint

doi: 10.64898/2026.03.30.715412

Figure Lengend Snippet: A, Schematic of glycolysis termination illustrating conditional Ldha:Ldhb:Pdha1:Slc16a1 deletion with Mx1Cre . B–C, MCT1 inhibition with AZD3965 in chimeric transplant recipients reduces the fraction of donor Ldha Δ/Δ ;Ldhb Δ/Δ ;Pdha1 Δ HSCs, progenitors, and mature cells: paired donor chimerism of B myeloid, B cells and C BM HSPCs (n = 10 mice from 3 independent experiments). D–E, Competitive bone marrow transplantation of donor CD45.2 + Mx1Cre;Slc16a1 Δ/Δ , Mx1Cre;Ldha Δ/Δ ;Ldhb Δ/Δ ; Slc16a1 Δ/Δ , or Ldha fl/fl ;Ldhb fl/fl ; Slc16a1 fl/fl littermate control bone marrow cells with wild-type CD45.1;CD45.2 competitor cells to lethally irradiated CD45.1 recipient mice. Shown are donor chimerism in D peripheral blood at weeks 4-16 and E BM HSPCs at week 16 (n = 11–15 mice per genotype for blood; n = 12–15 for BM; three independent experiments). F–G, Competitive transplantation of Mx1Cre;Ldha Δ/Δ ;Ldhb Δ/Δ ; Pdha1 Δ ; Slc16a1 Δ/Δ or littermate control BM cells. Donor contribution to F peripheral blood at weeks 4-16 and G BM HSPCs at week 16 (n = 13 for blood; n = 12–13 for BM; three independent experiments). Data represent mean ± s.d. Statistical significance was assessed with a paired t-test ( B ), one-way ANOVA ( C, D - CD45 + , Myeloid, B cells, E ), a Brown-Forsythe ANOVA test ( D : T cells), t-test ( F – Myeloid cells), Mann-Whitney test ( F- CD45 + , B, T cells) and Welch’s test ( G ). All figures show * p<0.05, ** p<0.01, ***p<0.001.

Article Snippet: For competitive transplantation experiments, 500,000-1,000,000 cells each of donor (CD45.2) and competitor (CD45.1/CD45.2) cells were mixed and transplanted into the retro-orbital venous sinus of anesthetized CD45.1 recipients irradiated using an XRAD 320 X-ray irradiator (Precision X-Ray) with two doses of 540 rad (1080 rad in total) at least 3 hours apart.

Techniques: Inhibition, Transplantation Assay, Control, Irradiation, MANN-WHITNEY