Journal: Frontiers in Molecular Neuroscience

Article Title: Cohesive Regulation of Neural Progenitor Development by microRNA miR-26, Its Host Gene Ctdsp and Target Gene Emx2 in the Mouse Embryonic Cerebral Cortex

doi: 10.3389/fnmol.2018.00044

Figure Lengend Snippet: Ctdsp2 increases neural progenitor (NP) proliferation. (A–C) Overexpression of Ctdsp2 , but not the control construct pCAGIG, increased the proportion of cells expressing both proliferative marker bromodeoxyuridine + (BrdU + )/green fluorescence protein + (GFP + ) and radial glial cell (RGC) marker Pax6 + /GFP + , but not intermediate progenitor (IP) marker Tbr2 + /GFP + , in GFP-positive cells. (D–F) shRNA-mediated knockdown (sh Ctdsp2 ) of Ctdsp2 decreased the proportion of both BrdU + /GFP + cells and Pax6 + /GFP + cells, but not Tbr2 + /GFP + cells in GFP-positive cells, compared to the control construct pSilencer. Values represent mean ± SEM. n = 9 sections from at least three brains. ** P < 0.01; ns, not significant. ANOVA with post hoc test was used. Scale bar = 50 μm.

Article Snippet: Primary antibodies against the following antigens were used: BrdU (1:50, Developmental Studies Hybridoma Bank at University of Iowa (DSHB)), Ki67 (1:500, Abcam), Pax6 (1:500, Covance), Pax6 (1:15 DSHB), Tbr2 (1:500, Abcam), GFP (1:1000, Abcam, chicken), and GFP (1:1000, Rockland, rabbit).

Techniques: Over Expression, Control, Construct, Expressing, Marker, Fluorescence, shRNA, Knockdown

Journal: Frontiers in Molecular Neuroscience

Article Title: Cohesive Regulation of Neural Progenitor Development by microRNA miR-26, Its Host Gene Ctdsp and Target Gene Emx2 in the Mouse Embryonic Cerebral Cortex

doi: 10.3389/fnmol.2018.00044

Figure Lengend Snippet: miR-26 promotes NP proliferation. (A,D,G) Overexpression of miR-26a, but not the control construct pCAGIG, proportion of cells expressing both proliferative marker BrdU + /GFP + and RGC marker Pax6 + /GFP + , but not IP marker Tbr2 + /GFP + , in GFP-positive cells. (B,E,H) miRNA sponge-mediated knockdown (miR-26-SP), but not the mutated sponge (miR-26-SPmut), decreased proportion of cells expressing both proliferative marker BrdU + /GFP + and RGC marker Pax6 + /GFP + in GFP-positive cells. (C,F,I) Ratio of BrdU + /GFP + , Pax6 + /GFP + or Tbr2 + /GFP + cells vs. GFP + cells in the electroporated cortex. Values represent mean ± SEM. n = 9 sections from at least three brains. * P < 0.05; ** P < 0.01; ns, not significant. ANOVA with post hoc test was used. Scale bar = 50 μm.

Article Snippet: Primary antibodies against the following antigens were used: BrdU (1:50, Developmental Studies Hybridoma Bank at University of Iowa (DSHB)), Ki67 (1:500, Abcam), Pax6 (1:500, Covance), Pax6 (1:15 DSHB), Tbr2 (1:500, Abcam), GFP (1:1000, Abcam, chicken), and GFP (1:1000, Rockland, rabbit).

Techniques: Over Expression, Control, Construct, Expressing, Marker, Knockdown

Journal: Frontiers in Molecular Neuroscience

Article Title: Cohesive Regulation of Neural Progenitor Development by microRNA miR-26, Its Host Gene Ctdsp and Target Gene Emx2 in the Mouse Embryonic Cerebral Cortex

doi: 10.3389/fnmol.2018.00044

Figure Lengend Snippet: Emx2 is functionally inhibited by miR-26 in regulating NP proliferation. (A–F) Overexpression of Emx2 , but not the control construct pCAGIG, decreased the proportion of cells expressing both proliferative marker BrdU + /GFP + and RGC marker Pax6 + /GFP + , but not IP marker Tbr2 + /GFP + , in GFP-positive cells. short hairpin RNA (shRNA)-mediated knockdown (sh Emx2 ) of Emx2 increased the proportion of both BrdU + /GFP + cells and Pax6 + /GFP + cells, but not Tbr2 + /GFP + cells in GFP-positive cells, compared to the control construct pSilencer. (G–J) Emx2 expression suppressed the proportion of cells expressing both proliferative marker BrdU + /GFP + and RGC marker Pax6 + /GFP + in GFP-positive cells. Co-expressing Emx2 with miR-26, but not miR-26-mut, dramatically reversed the suppression. (K,L) Emx2 expression did not alter the proportion of cells expressing IP marker Tbr2 + /GFP + , in GFP-positive cells. Values represent mean ± SEM. n = 9 sections from at least three brains. ** P < 0.01; *** P < 0.001; ns, not significant. ANOVA with post hoc test was used. Scale bar = 50 μm.

Article Snippet: Primary antibodies against the following antigens were used: BrdU (1:50, Developmental Studies Hybridoma Bank at University of Iowa (DSHB)), Ki67 (1:500, Abcam), Pax6 (1:500, Covance), Pax6 (1:15 DSHB), Tbr2 (1:500, Abcam), GFP (1:1000, Abcam, chicken), and GFP (1:1000, Rockland, rabbit).

Techniques: Over Expression, Control, Construct, Expressing, Marker, shRNA, Knockdown

Journal: Frontiers in Immunology

Article Title: 3-Hydroxystearic acid promotes cholesterol efflux and attenuates atherosclerosis via the ALKBH5/PAX-8/ABCA1 pathway

doi: 10.3389/fimmu.2026.1750021

Figure Lengend Snippet: C18-3OH modulates ABCA1 expression and cholesterol efflux in foam cells via PAX-8 upregulation. (A) RT-qPCR analysis of PAX-8 mRNA expression in foam cells treated with C18-3OH. (B) Western blot analysis of PAX-8 protein expression in foam cells treated with C18-3OH. ** P < 0.01 vs. control, n = 3. (C) Western blot detection of PAX-8 and ABCA1 protein expression in foam cells following PAX-8 knockdown. (D) Quantitative analysis of PAX-8 protein expression (Panel C). (E) RT-qPCR analysis of ABCA1 mRNA expression in foam cells after PAX-8 knockdown. (F) Quantitative analysis of ABCA1 protein expression (Panel C). (G) Effects of C18-3OH and siPAX-8 on cholesterol efflux in foam cells. * P < 0.05, ** P < 0.01 vs. siNC + DMSO; ## P < 0.01 vs. siNC + C18-3OH, n = 3. (H) NBD-cholesterol fluorescence intensity in foam cells; scale bar = 150 μm.

Article Snippet: Primary antibodies diluted in antibody dilution buffer were applied as follows: ABCA1 (Abcam, 1:1000), PAX-8 (Proteintech, 1:3000), ALKBH5 (Proteintech, 1:5000), ELAVL1 (Proteintech, 1:5000); GAPDH (Proteintech, 1:20, 000) and α-Tubulin (Proteintech, 1:20, 000) served as loading controls.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Control, Knockdown, Fluorescence

Journal: Frontiers in Immunology

Article Title: 3-Hydroxystearic acid promotes cholesterol efflux and attenuates atherosclerosis via the ALKBH5/PAX-8/ABCA1 pathway

doi: 10.3389/fimmu.2026.1750021

Figure Lengend Snippet: PAX-8 binds to and regulates ABCA1 expression. (A) Predicted binding sites between PAX-8 and ABCA1 in humans. (B) Predicted binding sites between PAX-8 and ABCA1 in mice. (C) Western blot analysis of PAX-8 and ABCA1 protein expression in foam cells following PAX-8 overexpression. (D) Quantitative analysis of PAX-8 protein expression (Panel C). (E) RT-qPCR analysis of ABCA1 mRNA expression in foam cells after PAX-8 overexpression. (F) Quantitative analysis of ABCA1 protein expression (Panel C). ** P < 0.01 vs. LV-NC, n = 3. (G) ChIP-Seq identification of PAX-8 binding to the ABCA1 locus. Genomic tracks showing PAX-8 binding to the ABCA1 locus in the Input (negative control, top) and ChIP (PAX-8-enriched, bottom) groups. The y-axis indicates relative enrichment of ChIP-Seq signals. The x-axis represents genomic coordinates. (H) ChIP-qPCR validation of PAX-8 targeting regulation of ABCA1. ** P < 0.01 vs. Input, n = 3.

Article Snippet: Primary antibodies diluted in antibody dilution buffer were applied as follows: ABCA1 (Abcam, 1:1000), PAX-8 (Proteintech, 1:3000), ALKBH5 (Proteintech, 1:5000), ELAVL1 (Proteintech, 1:5000); GAPDH (Proteintech, 1:20, 000) and α-Tubulin (Proteintech, 1:20, 000) served as loading controls.

Techniques: Expressing, Binding Assay, Western Blot, Over Expression, Quantitative RT-PCR, ChIP-sequencing, Negative Control, ChIP-qPCR, Biomarker Discovery

Journal: Frontiers in Immunology

Article Title: 3-Hydroxystearic acid promotes cholesterol efflux and attenuates atherosclerosis via the ALKBH5/PAX-8/ABCA1 pathway

doi: 10.3389/fimmu.2026.1750021

Figure Lengend Snippet: Effects of C18-3OH on m 6 A modification of PAX-8 mRNA in foam cells. (A) Predicted score distribution of the human PAX-8 gene query sequence. (B) Predicted score distribution of the murine PAX-8 gene query sequence. (C) Partitioned statistical plot of predicted scores for the human PAX-8 gene query sequence. (D) Partitioned statistical plot of predicted scores for the murine PAX-8 gene query sequence. (E) MeRIP-qPCR analysis of the effect of C18-3OH on m 6 A modification of PAX-8 mRNA in foam cells. ** P < 0.01 vs. control, n = 3.

Article Snippet: Primary antibodies diluted in antibody dilution buffer were applied as follows: ABCA1 (Abcam, 1:1000), PAX-8 (Proteintech, 1:3000), ALKBH5 (Proteintech, 1:5000), ELAVL1 (Proteintech, 1:5000); GAPDH (Proteintech, 1:20, 000) and α-Tubulin (Proteintech, 1:20, 000) served as loading controls.

Techniques: Modification, Sequencing, Control

Journal: Frontiers in Immunology

Article Title: 3-Hydroxystearic acid promotes cholesterol efflux and attenuates atherosclerosis via the ALKBH5/PAX-8/ABCA1 pathway

doi: 10.3389/fimmu.2026.1750021

Figure Lengend Snippet: Effects of C18-3OH on ALKBH5 expression in foam cells. (A) Bioinformatic analysis of methylation-associated proteins regulating PAX-8 in THP-1 cells. (B) RT-qPCR analysis of ALKBH5 mRNA expression in foam cells treated with C18-3OH. (C) Western blot analysis of ALKBH5 protein expression in foam cells treated with C18-3OH. (D) RT-qPCR analysis of ELAVL1 mRNA expression in foam cells treated with C18-3OH. (E) Western blot analysis of ELAVL1 protein expression in foam cells treated with C18-3OH. * P < 0.05, ** P < 0.01 vs. control, n = 3.

Article Snippet: Primary antibodies diluted in antibody dilution buffer were applied as follows: ABCA1 (Abcam, 1:1000), PAX-8 (Proteintech, 1:3000), ALKBH5 (Proteintech, 1:5000), ELAVL1 (Proteintech, 1:5000); GAPDH (Proteintech, 1:20, 000) and α-Tubulin (Proteintech, 1:20, 000) served as loading controls.

Techniques: Expressing, Methylation, Quantitative RT-PCR, Western Blot, Control

Journal: Frontiers in Immunology

Article Title: 3-Hydroxystearic acid promotes cholesterol efflux and attenuates atherosclerosis via the ALKBH5/PAX-8/ABCA1 pathway

doi: 10.3389/fimmu.2026.1750021

Figure Lengend Snippet: ALKBH5 mediates C18-3OH regulation of PAX-8 and ABCA1 expression, PAX-8 mRNA m 6 A modification, and cholesterol efflux. (A) Western blot analysis of ALKBH5 expression in foam cells following lentiviral transfection for ALKBH5 overexpression. ** P < 0.01 vs. control, n = 3. (B) Western blot analysis of PAX-8 and ABCA1 protein expression in foam cells following ALKBH5 overexpression. (C) RT-qPCR analysis of ABCA1 mRNA expression in foam cells after ALKBH5 overexpression. (D) Quantitative analysis of ABCA1 protein expression (Panel B). (E) RT-qPCR analysis of PAX-8 mRNA expression in foam cells after ALKBH5 overexpression. (F) Quantitative analysis of PAX-8 protein expression (Panel B). (G) MeRIP-qPCR analysis of PAX-8 mRNA m6A modification in foam cells following ALKBH5 overexpression. (H) Effect of C18-3OH and LV-ALKBH5 on cholesterol efflux in foam cells. (I) NBD-cholesterol fluorescence intensity in foam cells; scale bar = 150 μm. * P < 0.05, ** P < 0.01 vs. LV-NC + DMSO; ## P < 0.01 vs. LV-NC + C18-3OH, n = 3.

Article Snippet: Primary antibodies diluted in antibody dilution buffer were applied as follows: ABCA1 (Abcam, 1:1000), PAX-8 (Proteintech, 1:3000), ALKBH5 (Proteintech, 1:5000), ELAVL1 (Proteintech, 1:5000); GAPDH (Proteintech, 1:20, 000) and α-Tubulin (Proteintech, 1:20, 000) served as loading controls.

Techniques: Expressing, Modification, Western Blot, Transfection, Over Expression, Control, Quantitative RT-PCR, Fluorescence

Journal: Frontiers in Immunology

Article Title: 3-Hydroxystearic acid promotes cholesterol efflux and attenuates atherosclerosis via the ALKBH5/PAX-8/ABCA1 pathway

doi: 10.3389/fimmu.2026.1750021

Figure Lengend Snippet: Effects of C18-3OH on ABCA1, PAX-8, ALKBH5 expression, and PAX-8 mRNA m 6 A methylation in Aortas of apoE −/− Mice. (A) Western blot analysis of ABCA1, PAX-8, and ALKBH5 protein expression in aortas of HFD-fed apoE −/− mice treated with C18-3OH and/or ALKBH5 overexpression. (B) RT-qPCR analysis of ABCA1 mRNA expression in aortas of apoE −/− mice. (C) RT-qPCR analysis of PAX-8 mRNA expression in aortas of apoE −/− mice. (D) RT-qPCR analysis of ALKBH5 mRNA expression in aortas of apoE −/− mice. (E) Quantitative analysis of ABCA1 protein expression (Panel A). (F) Quantitative analysis of PAX-8 protein expression (Panel A). (G) Quantitative analysis of ALKBH5 protein expression (Panel A). (H) MeRIP-qPCR analysis of PAX-8 mRNA m 6 A methylation levels in aortas of apoE −/− mice across experimental groups. ** P < 0.01 vs. Control; # P < 0.05, ## P < 0.01 vs. C18-3OH + AAV-NC. n=6.

Article Snippet: Primary antibodies diluted in antibody dilution buffer were applied as follows: ABCA1 (Abcam, 1:1000), PAX-8 (Proteintech, 1:3000), ALKBH5 (Proteintech, 1:5000), ELAVL1 (Proteintech, 1:5000); GAPDH (Proteintech, 1:20, 000) and α-Tubulin (Proteintech, 1:20, 000) served as loading controls.

Techniques: Expressing, Methylation, Western Blot, Over Expression, Quantitative RT-PCR, Control