Journal: Journal of Translational Medicine

Article Title: Myeloid leukemia-derived galectin-1 downregulates CAR expression to hinder cytotoxicity of CAR T cells

doi: 10.1186/s12967-023-04832-x

Figure Lengend Snippet: Galectins are highly expressed in myeloid leukemia cells A , Heat maps showing suppressive genes differentially expressed between myeloid and lymphoid leukemia cell lines and between primary AML and ALL samples. Genes are those that are up or down-regulated more than two-fold, except galectin-9 in primary AML and ALL samples. AML and ALL primary samples had two biological replicates, respectively. B–E , Histograms showing the expression of galectin-1, galectin-3, galectin-12, and galectin-9 in AML and ALL cell lines; Histograms and the MFI of galectin-1, galectin-3,and galectin-12 in primary AML and ALL samples. Data shown are mean ± SEM of three experimental replicates

Article Snippet: We added NH 4 Cl (20 mM), galectin-1 specific inhibitor anginex (Jiangsu Jitai, 10 µM), galectin-3 specific inhibitor G3-C12 (MCE, 30 µM), recombinant galectin-1 protein (Acro, 50 µg/ml), recombinant galectin 3 protein (MCE, 20 µg/ml), recombinant galectin 12 protein (MCE, 10 µg/ml) and galectin 12 antibody (CUSABIO) to the coculture system.

Techniques: Expressing

Journal: Journal of Translational Medicine

Article Title: Myeloid leukemia-derived galectin-1 downregulates CAR expression to hinder cytotoxicity of CAR T cells

doi: 10.1186/s12967-023-04832-x

Figure Lengend Snippet: Myeloid leukemia cells use galectin-1 to block CAR T cells function A , The specific lysis after coculture with U937 CD33 cells at a 1:5 in the presence of recombinant galectin 3 protein (20 µg/ml) and galectin-3 specific inhibitor G3-C12 (30 µM). B , The specific lysis after coculture with U937 CD33 cells at a 1:1 in the presence of recombinant galectin 12 protein (10 µg/ml) and galectin 12 antibody. C , The specific lysis, the percentage of CD69 expression in CD33 CAR T cells, and the percentage of surface CAR expression in CD33 CAR T cells, after coculture with U937 CD33 cells at a 1:10 E:T ratio for 12 h, in the presence of 10 µM galectin-1 inhibitory peptide (ANGINEX). D , The specific lysis and the percentage of surface CAR expression in CD33 CAR T cells, after coculture with U937 CD33 cells at a 50 µg/ml recombinant galectin-1 protein. E , The specific lysis and the percentage of surface CAR expression in CD33 CAR T cells, after coculture with U937 CD33 cells at a 1:1 ratio for 12 h in the presence of 10 µM galectin-1 inhibitory peptide (ANGINEX). F, Total number of CD123, CD33,CD19 and CLL-1 CAR-T cells in the experimental group with 50 µg/ml recombinant galectin-1 protein and in the control group. G , Annexin V positivity in CD123, CD33, CD19 and CLL1 CAR-T cells from experimental and control groups using 50 µg/ml recombinant galectin-1 protein

Article Snippet: We added NH 4 Cl (20 mM), galectin-1 specific inhibitor anginex (Jiangsu Jitai, 10 µM), galectin-3 specific inhibitor G3-C12 (MCE, 30 µM), recombinant galectin-1 protein (Acro, 50 µg/ml), recombinant galectin 3 protein (MCE, 20 µg/ml), recombinant galectin 12 protein (MCE, 10 µg/ml) and galectin 12 antibody (CUSABIO) to the coculture system.

Techniques: Blocking Assay, Lysis, Recombinant, Expressing

Journal: bioRxiv

Article Title: Large pilin subunits provide distinct structural and mechanical properties for the Myxococcus xanthus type IV pilus

doi: 10.1101/2023.07.22.550172

Figure Lengend Snippet: A. Representative micrograph of negatively stained T4P Mx . Scale bar, 200nm. B. Representative cryo-EM micrograph of T4P Mx . Scale bar, 500nm. C. Comparison of cryo-EM reconstructions of T4P and ribbon models of pilins from M. xanthus (left), N. gonorrhoeae (center), and the 2-chain G. sulfurreducens (right), where the transparent surfaces are the cryo-EM density maps, and one pilin subunit is shown in red in each. D. Cross section of the cryo-EM reconstruction of the T4P Mx with a single PilA subunit shown in red. E. Placement of the T4P Mx structure into the central slice of the subtomogram average of the piliated M. xanthus T4PM revealed by cryo-electron tomography . The T4P, the outer membrane pore (OM pore), OM, and IM are indicated. Scale bar, 10nm. F. Complete T4P Mx structure composed of T4P Mx and a tip complex. The top PilA of the T4P Mx is shown in red and PilA subunits of T4P Mx below are shown in yellow, the AlphaFold-Multimer model containing the four minor pilins (FimU3, PilV3, PilW3, PilX3) and PilY1.3 is shown in the indicated colors. The complete T4P Mx structure was generated by superposing the top PilA (red) of the T4P Mx with the bottom PilA (wheat) of the AlphaFold-Multimer model (RMSD=0.841, Fig.S2E). The superposed PilA from the AlphaFold-Multimer model is not shown in the complete T4P Mx structure.

Article Snippet: The corresponding cryo-EM map was deposited in the EMDB with accession code EMD-41298.

Techniques: Staining, Cryo-EM Sample Prep, Tomography, Generated

Journal: Cancers

Article Title: Autocrine Signaling of NRP1 Ligand Galectin-1 Elicits Resistance to BRAF-Targeted Therapy in Melanoma Cells

doi: 10.3390/cancers12082218

Figure Lengend Snippet: Galectin-1 upregulation in melanoma samples and cell lines resistant to BRAF-targeted therapy. ( A ) The graph shows Galectin-1 (LGALS1 gene) mRNA expression variation in paired melanoma samples (obtained from seven patients, indicated by black diamonds) before treatment with BRAF inhibitors and after the onset of secondary resistance (Log2 ratio resistant/pretherapy), as well as in A375 and SK-MEL-28 melanoma cells before and after developing resistance to 2μM PLX-4720 (Log2 ratio Resistant/Parental; n = 3). ( B ) Analysis of Galectin-1 protein levels (by PikoKine ELISA assay) in conditioned media (CM) derived from either parental or PLX4720-resistant A375 and SK-MEL-28 (PLX-resistant) cells. Data are normalized to levels detected in the respective parental cells and represent the average of a technical duplicate. Statistical significance was assessed by t -test, comparing parental and their derived PLX-resistant cell lines; *** p < 0.0001.

Article Snippet: Galectin-1 concentrations in CM were measured by PikoKine ELISA assay (category EK0762; Boster Biological Technology, Pleasanton, CA, USA), according to the manufacturer’s protocol.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Derivative Assay

Journal:

Article Title: Xyloglucan Octasaccharide XXLGol Derived from the Seeds of Hymenaea courbaril Acts as a Signaling Molecule 1

doi:

Figure Lengend Snippet: Dose-response curves for α-l-fucosidase response induced by the xyloglucan oligomers XXLGoL (a), XXFGol (b), and XXXGol (d), the trisaccharide methyl α-l-Fuc-(1→2)-β-d-Gal(1→2)-β-d-Xyl (e), and the polymer (f). R. fruticosus protoplasts (2 × 106) in 25 mL of buffer were incubated for 15 min in the presence of inducer up to 100 nm. Each curve was obtained by least-squares regression of data from three to four replications carried out from three independent inducer sets. R is reported as the rate of α-l-fucosidase activity of treated over control protoplasts.

Article Snippet: Caylase 345 (cellulase) and Caylase M3 (pectinase) were purchased from Cayla (Toulouse, France). α-l-Fuc-(1→2)-β-d-Gal-(1→4)-β-d-Glc (2′-fucosyl-lactose) was from Oxford GlycoSystems (Coger, Paris, France).

Techniques: Incubation, Activity Assay

Journal:

Article Title: Xyloglucan Octasaccharide XXLGol Derived from the Seeds of Hymenaea courbaril Acts as a Signaling Molecule 1

doi:

Figure Lengend Snippet: Time course for α-l-fucosidase activation in R. fruticosus protoplasts challenged with the sugar inducers XXLGol (a), XXFGol (b), and XXXGol (d) and the trisaccharide methyl α-1-Fuc-(1→2)-β-d-Gal-(1→2)-β-d-Xyl (e). Protoplasts (2 × 106) in 25 mL of buffer were challenged with inducer used at the optimal concentration (0.1 nm XXXGol, 1 nm XXFGol, 5 nm XXLGol, and 10 nm trisaccharide). Each curve was obtained by least-squares regression of data from three to four replications carried out from three independent inducer sets. R is reported as the rate of α-l-fucosidase activity of treated over control protoplasts. Control sets were run without addition of 2,4-D or oligosaccharides.

Article Snippet: Caylase 345 (cellulase) and Caylase M3 (pectinase) were purchased from Cayla (Toulouse, France). α-l-Fuc-(1→2)-β-d-Gal-(1→4)-β-d-Glc (2′-fucosyl-lactose) was from Oxford GlycoSystems (Coger, Paris, France).

Techniques: Activation Assay, Concentration Assay, Activity Assay

Journal:

Article Title: Xyloglucan Octasaccharide XXLGol Derived from the Seeds of Hymenaea courbaril Acts as a Signaling Molecule 1

doi:

Figure Lengend Snippet: Effects of XXLGol (a), XXFGol (b), and XXXGol (d) and the trisaccharide methyl α-l-Fuc(1→2)-β-d-Gal(1→2)-β-d-Xyl (e) on 2,4-D-stimulated α-l-fucosidase in R. fruticosus protoplasts. The results are expressed as the R value in A and B and as the mean percentage of activation (inhibition) of 2,4-D-stimulated response in C. Protoplasts (2 × 106) in 25 mL of buffer were incubated for 15 min with sugar inducer up to 100 nm in the presence of 10 nm 2,4-D. Each curve was obtained by least-squares regression of data from three to four replications carried out from three independent inducer sets. The induced response given as R is reported as the rate of α-l-fucosidase activity of treated protoplasts over controls. The mean percentage of activation (inhibition) is calculated as R(2,4-D + oligosaccharide) − (R[2,4-D])/(R[2,4-D]) − R(control) × 100, where R(2,4-D + oligosaccharide) and R(2,4-D) are R values in protoplasts incubated with sugar inducer and 2,4-D and with 2,4-D, respectively, and R(control) is the R value in protoplasts suspended in buffer without the addition of 2,4-D or oligosaccharides.

Article Snippet: Caylase 345 (cellulase) and Caylase M3 (pectinase) were purchased from Cayla (Toulouse, France). α-l-Fuc-(1→2)-β-d-Gal-(1→4)-β-d-Glc (2′-fucosyl-lactose) was from Oxford GlycoSystems (Coger, Paris, France).

Techniques: Activation Assay, Inhibition, Incubation, Activity Assay

Journal:

Article Title: Xyloglucan Octasaccharide XXLGol Derived from the Seeds of Hymenaea courbaril Acts as a Signaling Molecule 1

doi:

Figure Lengend Snippet: Effects of XXLGol (a), XXFGol (b), XXXGol (d) and the trisaccharide methyl α-l-Fuc-(1→2)-β-d-Gal-(1→2)-β-d-Xyl (e) on GA3-stimulated α-l-fucosidase in R. fruticosus protoplasts. The results are expressed as the R value in A and B and as the mean percentage of activation of GA3-stimulated response in C. Protoplasts (2 × 106) in 25 mL of buffer were incubated for 15 min with sugar up to 100 nm in the presence of 10 nm GA3. Each curve was obtained by least-squares regression of data from three to four replications carried out from three independent inducer sets. R is reported as the rate of α-1-fucosidase activity of treated protoplasts over controls. The mean percentage of activation is calculated as = R(GA3 + oligosaccharide) − (R[GA3])/(R[GA3]) − R(control) × 100, where R(GA3 + oligosaccharide) and R(GA3) are R values in protoplasts incubated with sugar and GA3 and with GA3, respectively, and R(control) is the R value in protoplasts suspended in buffer without the addition of 2,4-D or oligosaccharides.

Article Snippet: Caylase 345 (cellulase) and Caylase M3 (pectinase) were purchased from Cayla (Toulouse, France). α-l-Fuc-(1→2)-β-d-Gal-(1→4)-β-d-Glc (2′-fucosyl-lactose) was from Oxford GlycoSystems (Coger, Paris, France).

Techniques: Activation Assay, Incubation, Activity Assay