Journal: Molecular Therapy. Nucleic Acids

Article Title: RNA activation of CEBPA improves leukemia treatment

doi: 10.1016/j.omtn.2025.102611

Figure Lengend Snippet: NOV340 liposomes deliver RNA into AML cells in vitro via macropinocytosis (A) Graphic of Cy3-MTL-CEBPA and controls. (B) Delivery of fluorescently tagged CEBPA-51 saRNA in myeloid leukemia and lymphoid leukemia cell lines. Two-way ANOVA with Holm-Sidak multiple comparison test; n = 3. Data represented as mean (SD). (C) Schematic of endocytosis inhibitors used for in vitro uptake inhibition assay. (D) Representative histograms of Cy3 fluorescence in AML cell lines after treatment with clathrin-mediated endocytosis inhibitor, Pitstop 2, or macropinocytosis inhibitors, EIPA or imipramine. (E) Changes in uptake of Cy3-MTL-CEBPA following treatment with endocytosis inhibitors in THP-1, MOLM-14, and KG1a cells. Two-way ANOVA with Holm-Sidak multiple comparison test; n = 3. Data represented as mean (SD).

Article Snippet: THP-1, HL-60, K562, and KG1a cell lines were obtained from the American Type Culture Collection (ATCC).

Techniques: Liposomes, In Vitro, Comparison, Inhibition, Fluorescence

Journal: Molecular Therapy. Nucleic Acids

Article Title: RNA activation of CEBPA improves leukemia treatment

doi: 10.1016/j.omtn.2025.102611

Figure Lengend Snippet: MTL-CEBPA upregulates CEBPA in AML in vitro models (A) Changes in CEBPA mRNA expression in THP-1, KG1a, or MOLM-14 cell lines 72-h post-MTL-CEBPA or control (MTL-FLUC) treatment. Two-tailed unpaired student t tests; n = 3. Data represented as mean (SD). (B) Representative western blots of THP-1 and MOLM-14 cell lines 96 h post-treatment. (C) Upregulation of CEBPA mRNA in ex vivo AML3 patient sample 48 h post MTL-CEBPA or control (MTL-FLUC) treatment. Two-tailed unpaired student t test; n = 3. Data represented as mean (SD).

Article Snippet: THP-1, HL-60, K562, and KG1a cell lines were obtained from the American Type Culture Collection (ATCC).

Techniques: In Vitro, Expressing, Control, Two Tailed Test, Western Blot, Ex Vivo

Journal: bioRxiv

Article Title: Combined TIRF and 3D Super-Resolution Microscopy for Nanoscopic Spatiotemporal Characterization of Adhesion Molecules on Microvilli

doi: 10.1101/2025.08.26.672391

Figure Lengend Snippet: Surface topography of KG1a cells reconstructed by TIRF microscopy. (a, b) Fluorescence images of KG1a cells stained by (a) MemBrite FX640 and (b) DiD-Vybrant membrane stains, captured using wide-field epi-illumination microscopy. (c) Fluorescence image of the bottom surface of the MemBrite FX640-stained KG1a cell, captured by TIRFM. (d) Filtered image of (c) using Laplacian of Gaussian (LoG) filter; Bottom view (top panel) and side view (bottom panel). The color bar indicates fluorescence intensity values. (e) 3D topographic map reconstructed from the fluorescence image shown in (d) using ; Bottom view (top panel) and side view (bottom panel). The color bar represents the distance from the glass surface. (f) Contours of individual microvilli in the topographic map shown in (e), obtained using an edge detection algorithm applied to the binary mask generated by thresholding the image. (g) Enlarged views of the regions highlighted by the rectangles in (f). δz values are red color-coded, with a step size of 10 nm. (h) SEM image of a KG1a cell, highlighting microvilli structures. (i) Comparative analysis of the average number of microvilli per µm² detected using SEM and TIRFM.

Article Snippet: KG1a cells, a human acute myelogenous leukemia cell line (ATCC), were maintained in RPMI (Gibco) supplemented with 10% fetal bovine serum, penicillin (100 U ml -1 ), and streptomycin (100 μg ml -1 ).

Techniques: Microscopy, Fluorescence, Staining, Membrane, Generated

Journal: bioRxiv

Article Title: Combined TIRF and 3D Super-Resolution Microscopy for Nanoscopic Spatiotemporal Characterization of Adhesion Molecules on Microvilli

doi: 10.1101/2025.08.26.672391

Figure Lengend Snippet: 3D coordinates of adhesion molecules on microvilli determined by 3D-SMLM. Panels (a–d) illustrate the data processing pipeline that filters and overlays SM localizations relative to microvilli structures. (a) 3D-SMLM localization map of CD44 on a KG1a cell, immunolabeled with AF-488-conjugated antibodies, captured at 0.6 μm z-axis depth. The color bar represents the z-axis position. The 3D-SMLM image was obtained from the same cell shown in . (b) Z-coordinate frequency distribution of individual localized CD44 molecules in (a). The cyan rectangle indicates the molecules adsorbed on the coverslip, whereas the blue rectangle highlights the molecules on the cell surface located within a 120 nm distance from the coverslip that were used for the following XY-filtration. (c) Binary mask image showing microvilli contour regions obtained from the 3D topographic map shown in with the threshold value of 200 counts. Mask applied to Z-filtered molecules, revealing that selected molecules localize to microvilli. (d) 3D overlay projection of the 3D topographic image obtained by TIRF microscopy displayed in the grayscale and the 3D coordinates of CD44 determined by 3D-SMLM and filtered by XY-filtration using the binary mask shown in red dots.

Article Snippet: KG1a cells, a human acute myelogenous leukemia cell line (ATCC), were maintained in RPMI (Gibco) supplemented with 10% fetal bovine serum, penicillin (100 U ml -1 ), and streptomycin (100 μg ml -1 ).

Techniques: Immunolabeling, Filtration, Microscopy

Journal: bioRxiv

Article Title: Combined TIRF and 3D Super-Resolution Microscopy for Nanoscopic Spatiotemporal Characterization of Adhesion Molecules on Microvilli

doi: 10.1101/2025.08.26.672391

Figure Lengend Snippet: Evaluation of the spatial superposition of the 3D cell surface topography and 3D coordinates of adhesion molecules. (a) 3D-SMLM localization map of actin on a KG1a cell labeled by AF-488-conjugated phalloidin and overlaid with a 3D cell surface topography image acquired by TIRFM with MemBrite FX640 stain. Blue dots indicate localized individual actin molecules, and the grayscale image shows the contours of microvilli on the cell surface. (b) 3D view of localized individual actin molecules superimposed on a single microvillus on a KG1a cell. (c, d) Bivariate frequency histograms showing the 3D distribution of actin molecules in the microvillus on a KG1a cell, projected along the (c) XZ and (d) YZ axes. (e) 3D view of the cell surface topography shown as a yellow mesh, overlaid with the spatial boundary obtained from the 3D coordinates of individual actin molecules shown as a cyan mesh. Blue dots indicate localized individual actin molecules. The axial distance from the coverslip surface is shown in grayscale. (f, g) Normalized 8-bit 2D images obtained from the (f) 3D topography of a single microvillus and (g) 3D spatial boundary obtained from the 3D coordinates of individual actin molecules. The axial distance from the coverslip surface is shown in grayscale. (h) Structural similarity (SSIM) index analysis of the 3D topography of the vertically- and horizontally-oriented microvilli generated by TIRFM and the 3D coordinates of the actin molecules determined by 3D-SMLM.

Article Snippet: KG1a cells, a human acute myelogenous leukemia cell line (ATCC), were maintained in RPMI (Gibco) supplemented with 10% fetal bovine serum, penicillin (100 U ml -1 ), and streptomycin (100 μg ml -1 ).

Techniques: Labeling, Staining, Generated

Journal: bioRxiv

Article Title: Combined TIRF and 3D Super-Resolution Microscopy for Nanoscopic Spatiotemporal Characterization of Adhesion Molecules on Microvilli

doi: 10.1101/2025.08.26.672391

Figure Lengend Snippet: Spatial distribution of adhesion molecules on microvilli. (a) 3D view of localized individual CD44 molecules immunolabeled with AF-488-conjugated antibodies, overlaid onto a single microvillus on a KG1a cell. (b, c) Bivariate frequency histograms showing the 3D distribution of CD44 molecules on the microvilli on a KG1a cell, projected along the (b) XZ and (c) YZ axes. (d) Structural similarity (SSIM) index analysis of the 3D topography of the vertically- and horizontally-oriented microvilli generated by TIRFM and the 3D coordinates of the CD44 molecules determined by 3D-SMLM. (e) 3D view of localized individual PSGL-1 molecules immunolabeled with AF-488-conjugated antibodies, overlaid onto a single microvillus on a KG1a cell. (f, g) Bivariate frequency histograms showing the 3D distribution of PSGL-1 molecules on the microvillus on a KG1a cell, projected along the (f) XZ and (g) YZ axes. (h) SSIM index analysis of the 3D topography of the vertically- and horizontally-oriented microvilli generated by TIRFM and the 3D coordinates of the PSGL-1 molecules determined by 3D-SMLM.

Article Snippet: KG1a cells, a human acute myelogenous leukemia cell line (ATCC), were maintained in RPMI (Gibco) supplemented with 10% fetal bovine serum, penicillin (100 U ml -1 ), and streptomycin (100 μg ml -1 ).

Techniques: Immunolabeling, Generated

Journal: bioRxiv

Article Title: Combined TIRF and 3D Super-Resolution Microscopy for Nanoscopic Spatiotemporal Characterization of Adhesion Molecules on Microvilli

doi: 10.1101/2025.08.26.672391

Figure Lengend Snippet: Effect of cell rolling on the nanoscopic 3D distribution of adhesion molecules relative to the 3D Microvilli topography. (a) Schematic illustration of the cell rolling assay using a microfluidic device. A suspension of KG1a cells was infused into the fluidic chamber coated with rh E-selectin using a syringe pump. (b) 3D view of localized individual actin molecules superimposed on a single microvillus on a KG1a cell rolled over E-selectin. (c, d) Bivariate frequency histograms showing the 3D distribution of actin molecules in the microvillus on a KG1a cell rolled over E-selectin, projected along the (c) XZ and (d) YZ axes. (e) 3D view of localized individual CD44 molecules immunolabeled with AF-488-conjugated antibodies, overlaid onto a single microvillus on a KG1a cell rolled over E-selectin. (f, g) Bivariate frequency histograms showing the 3D distribution of CD44 molecules on the microvilli on a KG1a cell rolled over E-selectin, projected along the (f) XZ and (g) YZ axes. (h) 3D view of localized individual PSGL-1 molecules immunolabeled with AF-488-conjugated antibodies, overlaid onto a single microvillus on a KG1a cell rolled over E-selectin. (i, j) Bivariate frequency histograms showing the 3D distribution of PSGL-1 molecules on the microvillus on a KG1a cell rolled over E-selectin, projected along the (i) XZ and (j) YZ axes.

Article Snippet: KG1a cells, a human acute myelogenous leukemia cell line (ATCC), were maintained in RPMI (Gibco) supplemented with 10% fetal bovine serum, penicillin (100 U ml -1 ), and streptomycin (100 μg ml -1 ).

Techniques: Suspension, Immunolabeling