biotin  (Bio-Rad)

Journal: bioRxiv

Article Title: Synergistic activation of TLR2 and Dectin-2 by mannose-capped lipoarabinomannan reprograms macrophage lipid metabolism in tuberculosis

doi: 10.64898/2026.02.18.706227

Figure Lengend Snippet: A. M. tuberculosis ManLAM (500 ng/ml), H 2 O 2 treated ManLAM (ML+H, 500 ng/ml), furfurman (FUR; commercial Dectin-2 agonist; 10 µg/ml) and vehicle control (C) were tested were tested for ability to induce NF-κB activation in HEK-Dectin-2 reporter cells. Data were generated and analyzed as in . B. Control and Dectin-2 knockdown (KD) iBMDM were treated 24h with 500ng/ml ManLAM preparations. Abbreviations are as in panel A. Imaging flow cytometry data were generated and expressed as in . C. Representative microscopy images (60x magnification) of vehicle-control (C) and treated iBMDM in the experiment shown in panel B. Panel description as in . D. iBMDM were treated with ManLAM (500 ng/ml) doses of furfurman (FUR; 2.5, 5 and 10 µg/ml). Imaging flow cytometry data were generated and expressed as in . E. Representative microscopy images (60x magnification) of vehicle-control (C) and treated iBMDM in the experiment shown in panel D. Panel description as in . *, p <0.05 by unpaired t test in all relevant panels.

Article Snippet: TLR2 ligand Pam3CSK4 and Dectin-2 ligand furfurman were purchased from Invivogen (San Diego, CA).

Techniques: Control, Activation Assay, Generated, Knockdown, Imaging, Flow Cytometry, Microscopy

Journal: bioRxiv

Article Title: Synergistic activation of TLR2 and Dectin-2 by mannose-capped lipoarabinomannan reprograms macrophage lipid metabolism in tuberculosis

doi: 10.64898/2026.02.18.706227

Figure Lengend Snippet: A. iBMDM were treated with Pam3CSK4 (PAM; 500 ng/ml) and furfurman (FUR; 10 µg/ml), singly or in combination (PAM+FUR), and corresponding vehicle controls. Imaging flow cytometry data were generated and expressed as in . B. Representative microscopy images (60x magnification) of vehicle-control (C) and treated iBMDM in the experiment shown in panel A. Panel description as in . C. M. tuberculosis ManLAM, demannosylated ManLAM (αtManLAM), and deacylated ManLAM (dManLAM) (all H 2 O 2 pre-treated, 500 ng/ml) and corresponding vehicle controls were tested for ability to induce NF-κB activation in HEK-TLR2 and HEK-Dectin 2 reporter cells. Treatment was for 24h as in preceding figures. Data are presented as ratio of OD 650 values obtained with treated vs vehicle control cells. The horizontal dotted line marks the corresponding vehicle-control reading. D. ManLAM and derivatives, as in panel C, were used to treat iBMDM (all 500 ng/ml for 24h). Imaging flow cytometry data were generated and expressed as in . E. Representative microscopy images (60x magnification) of vehicle-control (C) and treated iBMDM in the experiment shown in panel D. Panel description as in . *, p <0.05 by unpaired t test in all relevant panels.

Article Snippet: TLR2 ligand Pam3CSK4 and Dectin-2 ligand furfurman were purchased from Invivogen (San Diego, CA).

Techniques: Imaging, Flow Cytometry, Generated, Microscopy, Control, Activation Assay

Journal: bioRxiv

Article Title: Synergistic activation of TLR2 and Dectin-2 by mannose-capped lipoarabinomannan reprograms macrophage lipid metabolism in tuberculosis

doi: 10.64898/2026.02.18.706227

Figure Lengend Snippet: A. iBMDM, control (C, non-targeting guide RNAs), TLR2 knock-down (KD), and Dectin-2 (Dec-2) KD iBMDM clones were left uninfected or infected with M. tuberculosis mc 2 6206 (MOI= 15) for 48h in triplicate wells, washed, and stained with Bodipy 493/503 and antibodies to the macrophage marker F4/80 for imaging flow cytometry. Data were generated and expressed as in . B. Representative microscopy images (60x magnification) of uninfected (U) and infected (Mtb) iBMDM in the experiment shown in panel A. Panel description as in . C, D. iBMDM were treated with ManLAM (500ng/ml) and vehicle-treated or treated with increasing doses of the diacylglycerol transferase inhibitor, A922500 (DGAT-i; 60, 90, 120ng/ml) and the acyl-coenzyme A:cholesterol acyltransferase inhibitor, CAS 615264-52-3 (ACAT-i; 5, 10, 15µg/ml). Imaging flow cytometry data for lipid droplet content were generated and expressed as in . *, p <0.05 by unpaired t test in all relevant panels. E. iBMDM were treated with ManLAM (500ng/ml) and vehicle-treated or treated with increasing doses of the PPARγ inhibitor GW9662 (0.5, 1, 2µM), mTORC1 inhibitor rapamycin (0.2, 0.4, 0.8nM), and the NF-κB inhibitor JSH-23 (3.5, 7, 14µM). Imaging flow cytometry data for lipid droplet content were generated and expressed as in . *, p <0.05 by unpaired t test in all relevant panels.

Article Snippet: TLR2 ligand Pam3CSK4 and Dectin-2 ligand furfurman were purchased from Invivogen (San Diego, CA).

Techniques: Control, Knockdown, Clone Assay, Infection, Staining, Marker, Imaging, Flow Cytometry, Generated, Microscopy

Journal: bioRxiv

Article Title: Synergistic activation of TLR2 and Dectin-2 by mannose-capped lipoarabinomannan reprograms macrophage lipid metabolism in tuberculosis

doi: 10.64898/2026.02.18.706227

Figure Lengend Snippet: Acyl groups and mannose caps within ManLAM engage TLR2 and Dectin-2, respectively, activating an mTORC1-PPARγ pathway that promotes lipid droplet accumulation and an NF-κB pathway that induces inflammatory cytokine production (e.g., TNF-α). Lipid metabolic reprogramming proceeds largely independently of inflammatory signaling.

Article Snippet: TLR2 ligand Pam3CSK4 and Dectin-2 ligand furfurman were purchased from Invivogen (San Diego, CA).

Techniques: