Journal: Nature Plants

Article Title: Cell-type-specific autophagy in root-hair-forming cells is essential for salt stress tolerance in Arabidopsis thaliana

doi: 10.1038/s41477-026-02285-w

Figure Lengend Snippet: a . Confocal microscopy images of the trichoblasts and atrichoblasts in the root maturation zone of Arabidopsis wildtype Col-0 expressing ProUBQ10:GFP-ATG8A–I isoforms under nitrogen-starvation conditions. 5 days old Arabidopsis seedlings were incubated in nitrogen-deficient 1/2 MS media containing 2 μM concanamycin A for 2 h before imaging. Representative images of 5 replicates are shown. Area highlighted in the white-boxed region in the GFP-ATG8 + PI panel was further enlarged and presented in the inset panel. Scale bars, 30 μm. Inset scale bars, 10 μm. Green color, GFP-ATG8A–I isoforms. Magenta color, propidium iodide dye. T, trichoblasts. A, atrichoblasts. b . Quantification of the GFP-ATG8 puncta inside the vacuole per normalized area (10,000 μm 2 ) of the trichoblasts and atrichoblasts imaged in a . Bars indicate the mean ± SD of 5 replicates. Two-tailed and paired student t tests were performed to analyze the significance of GFP-ATG8 puncta density differences between the trichoblasts and the atrichoblasts. Exact p values are centered directly above the comparison brackets. c . Confocal microscopy images of trichoblasts and atrichoblasts of Arabidopsis wildtype Col-0 expressing ProATG8E:GFP-ATG8E. 5 days old Arabidopsis seedlings were incubated in 1/2 MS media containing 2 μM conA for 3 h before imaging. Representative images of 5 replicates are shown. Area highlighted in the white-boxed region in the GFP-ATG8E + PI panel was further enlarged and presented in the inset panel. Scale bars, 30 μm. Inset scale bars, 10 μm. Green color, GFP-ATG8E. Magenta color, propidium iodide dye. T, trichoblasts. A, atrichoblasts. d . Quantification of the GFP-ATG8E puncta inside the vacuole per normalized area (10,000 μm 2 ) of the trichoblasts and atrichoblasts imaged in c . Bars indicate the mean ± SD of 5 replicates. Two-tailed and paired student t test was performed to analyze the significance of GFP-ATG8E puncta density difference between the trichoblasts and the atrichoblasts. Exact p values are centered directly above the comparison brackets. e . Confocal microscopy images of epidermal cells in the root maturation zone of Col-0, atg5 , rhd6 rsl1 , cpc try , gl2 and wer myb23 expressing ProUBQ10:GFP-ATG8A under nitrogen-starvation conditions. 5-d-old Arabidopsis seedlings were incubated in nitrogen-deficient 1/2 MS media containing 2 μM concanamycin A for 2 h before imaging. Representative images of 5 replicates are shown. Area highlighted in the white-boxed region in the GFP-ATG8A + PI panel was further enlarged and presented in the inset panel. Scale bars, 30 μm. Inset scale bars, 10 μm. Green color, GFP-ATG8A. Magenta color, propidium iodide dye. A, the atrichoblast positions (adjacent to only one cortex cell). Note that the spindle-shaped ER bodies in wer myb23 (white arrow) were not inside the vacuole and were not counted as GFP-ATG8A puncta for autophagic flux quantification. f . Quantification of the GFP-ATG8A puncta inside the vacuole per normalized area (10,000 μm 2 ) of the cells at the trichoblast positions and the atrichoblast positions imaged in e . Bars indicate the mean ± SD of 5 replicates. Paired repeated measures one-way ANOVA and Fisher’s LSD tests were used to analyze the differences of the number of the GFP-ATG8A puncta between each group. Groups sharing the same letters (a/b/c) are not significantly different, while those with different letters are significantly different. Family-wise significance and confidence level, 0.05 (95% confidence interval).

Article Snippet: The coding sequence of Arabidopsis ATG5 (Supplementary Table ) was cloned into pTwist Amp High Copy to form the entry module pTwist-C- ATG5 -D (Twist Bioscience).

Techniques: Confocal Microscopy, Expressing, Incubation, Imaging, Two Tailed Test, Comparison

Journal: Nature Plants

Article Title: Cell-type-specific autophagy in root-hair-forming cells is essential for salt stress tolerance in Arabidopsis thaliana

doi: 10.1038/s41477-026-02285-w

Figure Lengend Snippet: a , Trichoblast–atrichoblast distribution patterns in Arabidopsis wild-type Col-0, the autophagy-defective mutant atg5 and the root-hair-development mutant lines rhd6 rsl1 , cpc try , gl2 and wer myb23 . b , c , Confocal microscopy images of epidermal cells in the root maturation zone of Col-0, atg5 , rhd6 rsl1 , cpc try , gl2 and wer myb23 expressing ProUBQ10 :: GFP-ATG8A under control ( b ) or NaCl ( c ) treatment. Five-day-old Arabidopsis seedlings were incubated in either control 1/2 MS media containing 2 μM conA for 2 h ( b ) or 1/2 MS media containing 50 mM NaCl + 1 μM conA for 40–60 min ( c ) before imaging. Representative images of five replicates are shown. The areas highlighted by white boxes in the GFP–ATG8A + PI panels are further enlarged and presented in the inset panels. Scale bars, 30 μm. Inset scale bars, 10 μm. Green indicates GFP–ATG8A; magenta indicates PI dye. T, trichoblast position (adjacent to two cortex cells); A, atrichoblast position (adjacent to only one cortex cell). Note that the spindle-shaped ER bodies in wer myb23 (white arrows in c ) were not inside the vacuole and were not counted as GFP–ATG8A puncta for autophagic flux quantification. d , e , Quantification of GFP–ATG8A puncta inside the vacuole per normalized area (10,000 μm 2 ) of the cells at the trichoblast positions and the atrichoblast positions imaged in b ( d ) or c ( e ). The bars indicate the mean ± s.d. of five replicates. Paired repeated-measures one-way analysis of variance (ANOVA) and Fisher’s least significant difference (LSD) tests were used to analyse the differences in the numbers of GFP–ATG8A puncta between each group. Groups sharing the same letters (a/b/c/d/e) are not significantly different, while those with different letters are significantly different. The family-wise significance and confidence level was 0.05 (95% confidence interval).

Article Snippet: The coding sequence of Arabidopsis ATG5 (Supplementary Table ) was cloned into pTwist Amp High Copy to form the entry module pTwist-C- ATG5 -D (Twist Bioscience).

Techniques: Mutagenesis, Confocal Microscopy, Expressing, Control, Incubation, Imaging

Journal: Nature Plants

Article Title: Cell-type-specific autophagy in root-hair-forming cells is essential for salt stress tolerance in Arabidopsis thaliana

doi: 10.1038/s41477-026-02285-w

Figure Lengend Snippet: a , Design of the trichoblast-specific ATG5 CRISPR mutagenesis plasmid expressed in wild-type Col-0 expressing ProUBQ10 :: mCherry-ATG8E ( mCherry-ATG8E in Col-0). b , Design of the trichoblast-specific ATG5 complementation plasmid transformed to complement the autophagy-defective mutant atg5 expressing ProUBQ10 :: mCherry-ATG8E ( mCherry-ATG8E in atg5 ). c , d , Confocal microscopy images of trichoblasts and atrichoblasts of Arabidopsis lines mCherry-ATG8E in Col-0, TSKO-R ( ATG5 mutagenized using ProRHD6 -driven Cas9), R-ATG5 (the atg5 mutant complemented with ProRHD6 -driven ATG5 ), E-ATG5 (the atg5 mutant complemented with ProEXP7 -driven ATG5 ) and mCherry-ATG8E in atg5 under control ( c ) or NaCl ( d ) treatment. Five-day-old Arabidopsis seedlings were incubated in either control 1/2 MS media containing 2 μM conA for 2 h ( c ) or 1/2 MS media containing 50 mM NaCl + 1 μM conA for 1 h ( d ) before imaging. Representative images of five replicates are shown. The areas highlighted by white boxes in the merge panels are further enlarged and presented in the inset panels. Scale bars, 30 μm. Inset scale bars, 10 μm. T, trichoblast; A, atrichoblast. e , f , Quantification of mCherry–ATG8E puncta inside the vacuole per normalized area (10,000 μm 2 ) of the trichoblasts and atrichoblasts imaged in c ( e ) or d ( f ). The bars indicate the mean ± s.d. of five replicates. Paired repeated-measures one-way ANOVA and Fisher’s LSD tests were used to analyse the differences in the numbers of mCherry–ATG8E puncta between each group. Groups sharing the same letters (a/b/c) are not significantly different, while those with different letters are significantly different. The family-wise significance and confidence level was 0.05 (95% confidence interval).

Article Snippet: The coding sequence of Arabidopsis ATG5 (Supplementary Table ) was cloned into pTwist Amp High Copy to form the entry module pTwist-C- ATG5 -D (Twist Bioscience).

Techniques: CRISPR, Mutagenesis, Plasmid Preparation, Expressing, Transformation Assay, Confocal Microscopy, Control, Incubation, Imaging

Journal: Nature Plants

Article Title: Cell-type-specific autophagy in root-hair-forming cells is essential for salt stress tolerance in Arabidopsis thaliana

doi: 10.1038/s41477-026-02285-w

Figure Lengend Snippet: a-d . RHD6 promoter is more efficient in disrupting autophagic flux in trichoblast cells compared to EXP7 promoter. a-b . Confocal microscopy images of trichoblasts and atrichoblasts in the root maturation zone of Arabidopsis lines mCherry-ATG8E in Col-0, TSKO-R ( ATG5 mutagenized using ProRHD6 -driven Cas9), TSKO-E ( ATG5 mutagenized using ProEXP7 -driven Cas9) and mCherry-ATG8E in atg5 under control ( a ) or ( b ) treatment. 5 days old Arabidopsis seedlings were incubated in either 1/2 MS media containing 2 μM concanamycin A for 2 h ( a ) or 1/2 MS media containing 50 mM NaCl + 1 μM concanamycin A for 1 h ( b ) before imaging. Representative images of 5 replicates are shown. Area highlighted in the white-boxed region in the merge panel was further enlarged and presented in the inset panel. Scale bars, 30 μm. Inset scale bars, 10 μm. T, trichoblasts. A, atrichoblasts. c-d . Quantification of the mCherry-ATG8E puncta inside the vacuole per normalized area (10,000 μm 2 ) of the cells of the trichoblasts and atrichoblasts imaged in a ( c ) or b ( d ). Bars indicate the mean ± SD of 5 replicates. Paired repeated measures one-way ANOVA and Fisher’s LSD tests were used to analyze the differences of the number of the mCherry-ATG8E puncta between each group. Groups sharing the same letters (a/b/c) are not significantly different, while those with different letters are significantly different. Family-wise significance and confidence level, 0.05 (95% confidence interval). e. Normal autophagosome structures can be observed in cells that do not express Cas9 in TSKO-R lines . Confocal microscopy images of cells of cotyledon epidermis and stele regions of TSKO-R. 5 days old Arabidopsis seedlings were incubated in control 1/2 MS media for 45 min before imaging. Representative images of 3 replicates are shown. Area highlighted in the white-boxed region in the merge panel was further enlarged and presented in the inset panel. Scale bars, 20 μm. Inset scale bars, 5 μm.

Article Snippet: The coding sequence of Arabidopsis ATG5 (Supplementary Table ) was cloned into pTwist Amp High Copy to form the entry module pTwist-C- ATG5 -D (Twist Bioscience).

Techniques: Confocal Microscopy, Control, Incubation, Imaging

Journal: Nature Plants

Article Title: Cell-type-specific autophagy in root-hair-forming cells is essential for salt stress tolerance in Arabidopsis thaliana

doi: 10.1038/s41477-026-02285-w

Figure Lengend Snippet: a . Confocal microscopy images showing sodium ion concentrations in the vacuoles of trichoblasts and atrichoblasts in the transition zone and the elongation zone of Arabidopsis wildtype Col-0 and the autophagy-defective mutants atg5 , atg8 and atg16 indicated by CoroNa Green AM staining. 5 days old Arabidopsis seedlings were incubated in control 1/2 MS media for 30 min and were subsequently incubated in control 1/2 MS media containing 2 μM CoroNa Green AM for 30 min before imaging. Representative images of 5 replicates are shown. Scale bars, 20 μm. Green color, CoroNa Green AM sodium indicator. Magenta color, propidium iodide dye. T, trichoblasts. A, atrichoblasts. b-c . Quantitative analysis of the vacuolar CoroNa Green AM fluorescence intensity ratio between trichoblasts and atrichoblasts at the transition zone ( b ) or elongation zone ( c ) of the Arabidopsis seedlings imaged in a . Bars indicate the mean ± SD of 5 replicates. Paired repeated measures one-way ANOVA and Fisher’s LSD tests were performed to analyze the significance of the CoroNa Green AM fluorescence intensity ratio differences between each group. Groups sharing the same letter ‘a’ are not significantly different. Family-wise significance and confidence level, 0.05 (95% confidence interval). d . Confocal microscopy images showing sodium ion concentrations in the vacuoles of trichoblasts and atrichoblasts in the transition zone and the elongation zone of Arabidopsis lines mCherry-ATG8E in Col-0, TSKO-R, R-ATG5, E-ATG5 and mCherry-ATG8E in atg5 indicated by CoroNa Green AM staining. 5 days old Arabidopsis seedlings were incubated in control 1/2 MS media for 30 min and were subsequently incubated in control 1/2 MS media containing 2 μM CoroNa Green AM for 30 min before imaging. Representative images of 5 replicates are shown. Scale bars, 20 μm. Green color, CoroNa Green AM sodium indicator (and the nuclear signals of Cas9-GFP in TSKO-R). Magenta color, mCherry-ATG8E. T, trichoblasts. A, atrichoblasts. e-f . Quantitative analysis of the vacuolar CoroNa Green AM fluorescence intensity ratio between trichoblasts and atrichoblasts in the transition zone ( e ) or elongation zone ( f ) of the Arabidopsis seedlings imaged in d . Bars indicate the mean ± SD of 5 replicates. Paired repeated measures one-way ANOVA and Fisher’s LSD tests were performed to analyze the significance of the CoroNa Green AM fluorescence intensity ratio differences between each group. Groups sharing the same letter ‘a’ are not significantly different. Family-wise significance and confidence level, 0.05 (95% confidence interval).

Article Snippet: The coding sequence of Arabidopsis ATG5 (Supplementary Table ) was cloned into pTwist Amp High Copy to form the entry module pTwist-C- ATG5 -D (Twist Bioscience).

Techniques: Confocal Microscopy, Staining, Incubation, Control, Imaging, Fluorescence

Journal: Nature Plants

Article Title: Cell-type-specific autophagy in root-hair-forming cells is essential for salt stress tolerance in Arabidopsis thaliana

doi: 10.1038/s41477-026-02285-w

Figure Lengend Snippet: a , Confocal microscopy images showing sodium ion accumulation in the vacuoles of trichoblasts and atrichoblasts in the root maturation zone of Arabidopsis wild-type Col-0 and the autophagy-defective mutants atg5 , atg8 and atg16 . Five-day-old Arabidopsis seedlings were incubated in control 1/2 MS media for 30 min and were subsequently incubated in control 1/2 MS media containing 2 μM CoroNa Green AM for 30 min before imaging. Representative images of five replicates are shown. The areas highlighted by white boxes in the CoroNa Green AM + PI panels are further enlarged and presented in the inset panels. Scale bars, 30 μm. Inset scale bars, 10 μm. Green indicates CoroNa Green AM sodium indicator; magenta indicates PI dye. T, trichoblast; A, atrichoblast. b , Quantitative analysis of the vacuolar CoroNa Green AM fluorescence intensity ratio between trichoblasts and atrichoblasts of the Arabidopsis seedlings imaged in a . The bars indicate the mean ± s.d. of five replicates. Paired repeated-measures one-way ANOVA and Fisher’s LSD tests were performed to analyse the significance of the CoroNa Green AM fluorescence intensity ratio differences between each group. Groups sharing the same letters (a/b) are not significantly different, while those with different letters are significantly different. The family-wise significance and confidence level was 0.05 (95% confidence interval). c , Phenotypic characterization of the seedlings of Arabidopsis wild-type Col-0 and the autophagy-defective mutants atg5 , atg8 and atg16 under NaCl stress treatment. Representative images of three replicates are shown. d , Quantitative analysis of the proportion of non-etiolated leaves of the Arabidopsis seedlings imaged in c . The bars indicate the mean ± s.d. of three replicates. Paired repeated-measures one-way ANOVA and Fisher’s LSD tests were performed to analyse the significance of the proportion differences between each group. Groups sharing the same letters (a/b) are not significantly different, while those with different letters are significantly different. The family-wise significance and confidence level was 0.05 (95% confidence interval). e , Confocal microscopy images showing the sodium ion concentrations in the vacuoles of trichoblasts and atrichoblasts in the root maturation zone of Arabidopsis lines mCherry-ATG8E in Col-0, TSKO-R, R-ATG5, E-ATG5 and mCherry-ATG8E in atg5 . Five-day-old Arabidopsis seedlings were incubated in control 1/2 MS media for 30 min and were subsequently incubated in control 1/2 MS media containing 2 μM CoroNa Green AM for 30 min before imaging. Representative images of five replicates are shown. The areas highlighted by white boxes in the CoroNa Green AM + mCherry–ATG8E panels are further enlarged and presented in the inset panels. Scale bars, 30 μm. Inset scale bars, 10 μm. Green indicates CoroNa Green AM sodium indicator (and the nuclear signals of Cas9–GFP in TSKO-R); magenta indicates mCherry–ATG8E. T, trichoblast; A, atrichoblast. f , Quantitative analysis of the vacuolar CoroNa Green AM fluorescence intensity ratio between trichoblasts and atrichoblasts of the Arabidopsis seedlings imaged in e . The bars indicate the mean ± s.d. of five replicates. Paired repeated-measures one-way ANOVA and Fisher’s LSD tests were performed to analyse the significance of the CoroNa Green AM fluorescence intensity ratio differences between each group. Groups sharing the same letters (a/b) are not significantly different, while those with different letters are significantly different. The family-wise significance and confidence level was 0.05 (95% confidence interval). g , Phenotypic characterization of the seedlings of Arabidopsis lines mCherry-ATG8E in Col-0, TSKO-R, R-ATG5, E-ATG5 and mCherry-ATG8E in atg5 under NaCl stress treatment. Representative images of ten replicates are shown. h , Quantitative analysis of the proportion of non-etiolated leaves of the Arabidopsis seedlings imaged in g . The bars indicate the mean ± s.d. of ten replicates. Paired repeated-measures one-way ANOVA and Fisher’s LSD tests were performed to analyse the significance of the proportion differences between each group. Groups sharing the same letters (a/b/c/d) are not significantly different, while those with different letters are significantly different. The family-wise significance and confidence level was 0.05 (95% confidence interval).

Article Snippet: The coding sequence of Arabidopsis ATG5 (Supplementary Table ) was cloned into pTwist Amp High Copy to form the entry module pTwist-C- ATG5 -D (Twist Bioscience).

Techniques: Confocal Microscopy, Incubation, Control, Imaging, Fluorescence

Journal: Nature Plants

Article Title: Cell-type-specific autophagy in root-hair-forming cells is essential for salt stress tolerance in Arabidopsis thaliana

doi: 10.1038/s41477-026-02285-w

Figure Lengend Snippet: a-b . Quantitative analysis of primary root length of Arabidopsis seedlings imaged in Fig. ( a , n = 24) or Fig. ( b , n = 70). Repeated measures one-way ANOVA and Tukey tests were performed to analyze the significance of the primary root elongation difference between each group. Groups sharing the same letter ‘a’ are not significantly different. Family-wise significance and confidence level, 0.05 (95% confidence interval). Black lines indicate the median. White lines, 25% and 75% quartiles. c-d . Quantitative analysis of the number of newly emerged lateral roots of Arabidopsis seedlings imaged in Fig. ( a , n = 24) or Fig. ( b , n = 70). Unpaired Brown-Forsythe and Welch ANOVA tests and two-sided unpaired t tests with Welch’s correction (for multiple comparison) were performed to analyze the significance of the number of newly emerged lateral roots between each group, n = 24. Groups sharing the same letters (a/b/c/d) are not significantly different, while those with different letters are significantly different. Family-wise significance and confidence level, 0.05 (95% confidence interval). Black lines indicate the median. White lines, 25% and 75% quartiles. Note that the median and 25% quartile lines of atg5 , atg8 , and atg16 were all at the base line (y = 0), and the median line of mCherry-ATG8E in atg5 , the 25% quartile lines of R-ATG5, E-ATG5 and mCherry-ATG8E in atg5 were all at the base line (y = 0).

Article Snippet: The coding sequence of Arabidopsis ATG5 (Supplementary Table ) was cloned into pTwist Amp High Copy to form the entry module pTwist-C- ATG5 -D (Twist Bioscience).

Techniques: Comparison

Journal: Nature Plants

Article Title: Cell-type-specific autophagy in root-hair-forming cells is essential for salt stress tolerance in Arabidopsis thaliana

doi: 10.1038/s41477-026-02285-w

Figure Lengend Snippet: a , Confocal microscopy images showing the relative ROS levels in the trichoblasts and atrichoblasts of Arabidopsis wild-type Col-0 and the autophagy-defective mutants atg5 , atg8 and atg16 . Five-day-old Arabidopsis seedlings were incubated in either 1/2 MS (control) or 1/2 MS + 50 mM NaCl (NaCl) liquid media for 45 min and were subsequently stained with 1 μM CM-H 2 DCFDA for 1 min before imaging. Representative images of five replicates are shown. The areas highlighted by white boxes in the CM-H 2 DCFDA panels are further enlarged and presented in the inset panels. Scale bars, 30 μm. Inset scale bars, 30 μm. T, trichoblast; A, atrichoblast. c , Confocal microscopy images showing the relative ROS levels in the trichoblasts and atrichoblasts of Arabidopsis lines mCherry-ATG8E in Col-0, TSKO-R, R-ATG5, E-ATG5 and mCherry-ATG8E in atg5 . Five-day-old Arabidopsis seedlings were incubated in either 1/2 MS (control) or 1/2 MS + 50 mM NaCl (NaCl) liquid media for 45 min and were subsequently stained with 1 μM CM-H 2 DCFDA for 1 min before imaging. Representative images of five replicates are shown. The areas highlighted by white boxes in the CM-H 2 DCFDA panels are further enlarged and presented in the inset panels. Scale bars, 30 μm. Inset scale bars, 30 μm. b , d , Quantitative analysis of the relative ROS ratio between trichoblasts and atrichoblasts of the Arabidopsis seedlings imaged in a ( b ) or c ( d ). The bars indicate the mean ± s.d. of five replicates. Paired repeated-measures two-way ANOVA and Fisher’s LSD tests were performed to analyse the significance of the CM-H 2 DCFDA fluorescence intensity ratio differences between each group. Groups sharing the same letter (a/b/c) are not significantly different, while those with different letters are significantly different. The family-wise significance and confidence level was 0.05 (95% confidence interval).

Article Snippet: The coding sequence of Arabidopsis ATG5 (Supplementary Table ) was cloned into pTwist Amp High Copy to form the entry module pTwist-C- ATG5 -D (Twist Bioscience).

Techniques: Confocal Microscopy, Incubation, Control, Staining, Imaging, Fluorescence

Journal: Nature Communications

Article Title: Multiplex gene-editing strategy to engineer allogeneic EGFR-targeting CAR T-cells with improved efficacy against solid tumors

doi: 10.1038/s41467-025-66737-1

Figure Lengend Snippet: a Flow cytometry staining of H226 tumor cells in vitro for CD73 expression. Isotype control (gray), anti-CD73 antibody (pink). b Immunofluorescent micrographs of H226 tumors resected from NCG mice 46 days post-implantation. Nucleated cells (DAPI, blue), hypoxia (Hypoxyprobe, green) and CD73 (pink). Representative images from four individual tumors from 10 to 20 different cutting surfaces. c Quantification of hypoxia in various tumor regions within resected H226 tumors from NCG mice determined by mean fluorescence intensity (MFI) of Hypoxyprobe. Representative image of a resected tumor section; quantification was performed across 6 independent slides (3 tumors per slide from individual mice) with an average of 13.5 regions of interest (ROI) analyzed per slide. d Spatial transcriptomics gene expression analysis from hypoxic regions in ( c ) (white = low hypoxia, light green = medium hypoxia, dark green = high hypoxia). Boxplots show the median (line), interquartile range (box), and whiskers extending to values within 1.5× the IQR. e 2 × 10 6 UTD T cells (white, n = 5 individual mice) or unedited (gray, n = 5 individual mice) and A 2A R-KO (red, n = 5 individual mice) CAR T-cells injected I.V. into H226 tumor-bearing NCG mice. Group average of tumor volumes measured via calipers over time (Two-sided Mann–Whitney t-test, n = group average of individual mice, mean ± SEM, P** = 0.0079, P** = 0.0072). f Cumulative tumor burden, calculated as area under the curve, from ( e ) (Two-sided Mann–Whitney t-test, n = average of individual mice as above, mean ± SD, n.s. = 0.0556, P** = 0.00379). g 2 × 10 6 UTD T cells (white, n = 5 individual mice) or unedited (gray, n = 5 individual mice) and A 2A R-KO (red, n = 5 individual mice) CAR T-cells injected I.V. into A549 tumor-bearing NCG mice. Group average of tumor volumes measured via calipers over time (Graph represents group mean ± SD, P** = 0.0072). h Cumulative tumor burden, calculated as area under the curve, from ( g ). (Two-sided Mann–Whitney t-test, n = average of individual mice as above, mean ± SD, n.s. = 0.490, P** = 0.0037). For all data, symbols and error bars reflect individual biological replicates and group mean ± S.E.M. e – h Mann–Whitney t-test performed to calculate statistical significance, ** P < 0.01, * P < 0.05.

Article Snippet: Representative image of a resected tumor section; quantification was performed across 6 independent slides (3 tumors per slide from individual mice) with an average of 13.5 regions of interest (ROI) analyzed per slide. d Spatial transcriptomics gene expression analysis from hypoxic regions in ( c ) (white = low hypoxia, light green = medium hypoxia, dark green = high hypoxia).

Techniques: Flow Cytometry, Staining, In Vitro, Expressing, Control, Fluorescence, Gene Expression, Injection, MANN-WHITNEY

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Diversity, expression, and structural modeling of sugar transporters in Anisakis simplex s. s. L3 and L4 larvae: an in vitro and in silico study

doi: 10.3389/fcimb.2025.1621051

Figure Lengend Snippet: The phylogeny tree of the glucose transporter family. The figure shows a phylogenetic tree of facilitative glucose transporters (FGTs) and the “Sugars Will Eventually Be Exported Transporters” (SWEETs) of vertebrates, nematodes and insects. A rooted tree was constructed with the neighbor-joining method using MEGA ver. 7. The numbers on the internal branches are the bootstrap values (only >50% are shown). The sequences are labeled with data from GenBank, such as species name, name and accession number of the sugar transporters. The glucose transporters of A. simplex are indicated by the red frame. The sucrose transporters of Drosophila melanogaster are highlighted in purple, the GLUTs of Homo sapiens in blue, the transporters of Toxocara canis in green and those of Caenorhabditis spp. in orange.

Article Snippet: The mRNA expression levels of the putative facilitated glucose transporter genes ( fgt-1 , fgt-2 , fgt-3 , fgt-5 , fgt-9 ) and one Sugars Will Eventually be Exported Transporter ( sweet-1 ) were determined in L3 and L4 stage larvae of A. simplex s. s. The quantitative real-time PCR was performed using a QuantStudio 3 System (Applied Biosystems, Foster City, CA, USA).

Techniques: Construct, Labeling