Journal: The FASEB Journal

Article Title: Dissecting Shared Genetic Architecture of Thoracic Aortic Aneurysm and Aortic Related Traits and Identifying SplA /Ryanodine Receptor Domain and SOCS Box Containing 1 Involved in Smooth Muscle Phenotype Switching and Cell Senescence Through Alternative Splicing

doi: 10.1096/fj.202502457R

Figure Lengend Snippet: SplA/Ryanodine Receptor Domain and SOCS Box Containing 1 increases in thoracic aortic aneurysm models and thoracic aortic aneurysm patients. (A) Immunostaining for SplA/Ryanodine Receptor Domain and SOCS Box Containing 1 (SPSB1) in thoracic aortic aneurysm (TAA) patients ( n = 7 in TAA group, n = in non‐TAA group). The bar plot show quantification of the percentage of SPSB1 positive areas; (B) Western blot analysis for SPSB1 in TAA patients, β‐tubulin was used as loading control ( n = 4/groups); (C) Ultrasonic plot of β‐aminopropionitrile (BAPN)‐induced thoracic aortic pathology models, transverse aortic constriction (TAC) models, and corresponding control groups in the phase of end diastole. The bar plot shows the mean lumen diameter of ascending aorta of each group ( n = 17 in BAPN group, n = 15 in BAPN‐control group, n = 16 in TAC group, n = 15 in TAC sham‐operated group). (D) Real‐time quantitative polymerase chain reaction (qPCR) was used to determine the expression of SPSB1 in BAPN‐induced thoracic aortic pathology models, TAC models, and corresponding control groups ( n = 6 in BAPN group, n = 5 in BAPN‐control group, n = 6 in TAC group, n = 6 in TAC sham‐operated group). (E) Hematoxylin and eosin staining (H&E), Sirius Red staining, and Immunostaining in the cross‐sections of thoracic aorta ( n = 5 in BAPN group, n = 3 in BAPN‐control group, n = 4 in TAC group, n = 4 in TAC sham‐operated group). Immunostaining for SPSB1 was conducted in BAPN‐induced thoracic aortic pathology models, TAC models, and corresponding control groups. The bar plot shows quantification of the percentage of SPSB1 positive areas. Statistical analysis was performed using two‐tailed Student's t ‐test. * p < 0.05; ** p < 0.01; *** p < 0.001; (F) Western blot analysis for SPSB1 in BAPN models and control group, β‐tubulin was used as loading control ( n = 8 in BAPN group, n = 7 in control group); (G) Western blot analysis for SPSB1 in TAC models and control group, β‐tubulin was used as loading control ( n = 6 in TAC group, n = 6 in sham‐operated group. In the same group of mice, tissue were randomly pooled in pairs for protein extraction). SiSPSB1, SPSB1 knockdown with small interfering RNAs; ctrl, control group; sham, sham‐operated group.

Article Snippet: Small interfering RNAs (siRNAs) targeting SPSB1 were synthesized by MCE Corporation (Guangzhou, China).

Techniques: Immunostaining, Western Blot, Control, Real-time Polymerase Chain Reaction, Expressing, Staining, Two Tailed Test, Protein Extraction, Knockdown

Journal: The FASEB Journal

Article Title: Dissecting Shared Genetic Architecture of Thoracic Aortic Aneurysm and Aortic Related Traits and Identifying SplA /Ryanodine Receptor Domain and SOCS Box Containing 1 Involved in Smooth Muscle Phenotype Switching and Cell Senescence Through Alternative Splicing

doi: 10.1096/fj.202502457R

Figure Lengend Snippet: SplA/Ryanodine Receptor Domain and SOCS Box Containing 1 regulates smooth muscle cell phenotype switching and cell senescence. (A) SplA/Ryanodine Receptor Domain and SOCS Box Containing 1 (SPSB1) expression among distinct cellular populations; (B) SPSB1 was knockdown in aortic smooth muscle cell (SMC). The mRNA levels of α‐SMA, SM22, and KLF4 were detected by Real‐time quantitative polymerase chain reaction (qPCR) ( n = 6/experiments); (C) Immunofluorescent staining for α‐SMA (green), SM22 (green), KLF4 (green), and staining with DAPI (blue) in the aortic SMC after SPSB1 silenced; (D) Western blot analysis of the indicated proteins in SPSB1 knockdown aortic SMCs. β‐tubulin was used as a loading control for Western blotting ( n = 4/experiments). (E) Representative images of SA‐β‐gal (senescence‐associated β‐galactosidase)–stained SPSB1 knockdown aortic SMC and statistical analysis (right) ( n = 7 in control group, n = 5 in siRNA group). The blue regions are positively stained. Scale bar = 100 μm.; (F) Western blot analysis of senescence markers in SPSB1 knockdown aortic SMCs. β‐tubulin was used as a loading control for Western blotting ( n = 4/experiments); (G) Quantification of mRNA levels of senescence‐associated secretory phenotype (SASP) components in SPSB1 knockdown aortic SMCs ( n = 6/experiments). Statistical analysis was performed using two‐tailed Student's t ‐test; (H) Interleukin 6 (IL‐6) concentration in the culture supernatant of SPSB1 knockdown aortic SMCs. Statistical analysis was performed using two‐tailed Student's t ‐test ( n = 4/experiments); (I) The activity of MMP2 and MMP9 in culture medium was measured by gel zymography in SPSB1 knockdown aortic SMCs. SiSPSB1, SPSB1 knockdown with small interfering RNAs; ctrl, control group.

Article Snippet: Small interfering RNAs (siRNAs) targeting SPSB1 were synthesized by MCE Corporation (Guangzhou, China).

Techniques: Expressing, Knockdown, Real-time Polymerase Chain Reaction, Staining, Western Blot, Control, Two Tailed Test, Concentration Assay, Activity Assay, Zymography

Journal: The FASEB Journal

Article Title: Dissecting Shared Genetic Architecture of Thoracic Aortic Aneurysm and Aortic Related Traits and Identifying SplA /Ryanodine Receptor Domain and SOCS Box Containing 1 Involved in Smooth Muscle Phenotype Switching and Cell Senescence Through Alternative Splicing

doi: 10.1096/fj.202502457R

Figure Lengend Snippet: SplA/Ryanodine Receptor Domain and SOCS Box containing 1 regulates global alternative pre‐mRNA splicing profiles in aortic smooth muscle cells. (A) Gene Ontology (GO) enrichment analysis of SplA/Ryanodine Receptor Domain and SOCS Box Containing 1 (SPSB1) co‐expressed genes in contractile smooth muscle cells (SMC) in single‐cell RNA‐sequencing; (B) GO enrichment analysis of quantitative proteomics in SPSB1 knockdown aortic SMCs; (C) Heatmap showing percent spliced‐in (PSI) values for differentially spliced alternative splicing events (DAS) between SPSB1 knockdown aortic SMCs and control group ( n = 3/group); (D) Dot plot showing the distribution of PSI values for five AS types upon SPSB1 knockdown in aortic SMCs; (E) Bar plot of the DAS observed in RNA‐seq analysis of SPSB1 knockdown aortic SMCs and control group; (F) Schematic (right) and distribution (left) of five alternative splicing event types that differentially change upon SPSB1 knockdown aortic SMCs; (G) Representative Sashimi plot showing the decreased SEs of the RPL17, ATM, FOXM1, and MRPL43 genes in SPSB1 knockdown aortic SMCs and control group. SiSPSB1, SPSB1 knockdown with small interfering RNAs; ctrl, control group; SE, skipped exon; RI, retained intron; A3SS, alternative 3′ splice site; A5SS, alternative 5′ splice site; MXE, mutually exclusive exon.

Article Snippet: Small interfering RNAs (siRNAs) targeting SPSB1 were synthesized by MCE Corporation (Guangzhou, China).

Techniques: RNA Sequencing, Quantitative Proteomics, Knockdown, Alternative Splicing, Control

Journal: International Dental Journal

Article Title: Forkhead Box O1 Promotes Osteogenic Differentiation of Periodontal Ligament Stem Cells in Hypoxia/Reoxygenation Environments by Regulating Heme Oxygenase-1 Scavenging of Reactive Oxygen Species

doi: 10.1016/j.identj.2025.100894

Figure Lengend Snippet: H/R reduced cell viability, enhanced ROS accumulation, increased HIF-1α expression, and attenuated FoxO1 expression in PDLSCs. (A) The proliferation ability of PDLSCs was detected using CCK-8. (B) mRNA expression of HIF-1α in the normoxic-OM and H/R-OM groups. (C) Representative images of 2′,7′-dichlorodihydrofluorescein diacetate (DCFH-DA) staining in the normoxic control group and the H/R group. (D) Quantitative analysis of ROS. (E) The mRNA expression level of FoxO1 in the normoxic-control, normoxic-OM, H/R-control, and H/R-OM groups. * P < .05; ** P < .01; *** P < .001; **** P < .0001.

Article Snippet: Small-interfering RNA (siRNA) and lentivirus targeting human FoxO1 were purchased from GenePharma.

Techniques: Expressing, CCK-8 Assay, Staining, Control

Journal: International Dental Journal

Article Title: Forkhead Box O1 Promotes Osteogenic Differentiation of Periodontal Ligament Stem Cells in Hypoxia/Reoxygenation Environments by Regulating Heme Oxygenase-1 Scavenging of Reactive Oxygen Species

doi: 10.1016/j.identj.2025.100894

Figure Lengend Snippet: FoxO1 overexpression attenuated the inhibitory effect of H/R on the osteogenic differentiation of PDLSCs. (A) The efficacy of FoxO1 inhibition by siRNA was measured based on RT-qPCR. (B) Transfection with different multiplicity of infection (MOI) of Len-FoxO1 in PDLSCs. (C) Transcript level of FoxO1. (D) mRNA expression level of osteogenesis markers, including COL-I, RunX2, and ALP, in normoxic-OM, H/R-OM, H/R-OM + si-FoxO1 and H/R-OM + Len-FoxO1 groups. (E) ALP staining after osteogenic induction for 7 days. (F) ALP activity assay after 7 days of osteogenic induction. (G) ARS staining after osteogenic induction for 14 days. (H) Quantification of calcium deposits after 14 days of osteogenic induction. (I) Detection of intracellular ROS levels in the normoxic-OM, H/R-OM, and H/R-OM + si-FoxO1 groups. (J) Quantitative analysis of ROS. * P < .05; ** P < .01; *** P < .001; **** P < .0001.

Article Snippet: Small-interfering RNA (siRNA) and lentivirus targeting human FoxO1 were purchased from GenePharma.

Techniques: Over Expression, Inhibition, Quantitative RT-PCR, Transfection, Infection, Expressing, Staining, ALP Activity Assay

Journal: International Dental Journal

Article Title: Forkhead Box O1 Promotes Osteogenic Differentiation of Periodontal Ligament Stem Cells in Hypoxia/Reoxygenation Environments by Regulating Heme Oxygenase-1 Scavenging of Reactive Oxygen Species

doi: 10.1016/j.identj.2025.100894

Figure Lengend Snippet: mRNA levels of HO-1 in the normoxic-OM, H/R-OM, H/R-OM + si-FoxO1 and H/R-OM + Len-FoxO1 groups. * P < .05; ** P < .01; *** P < .001; **** P < .0001.

Article Snippet: Small-interfering RNA (siRNA) and lentivirus targeting human FoxO1 were purchased from GenePharma.

Techniques:

Journal: Redox Biology

Article Title: Divergent redox responses of macular and peripheral Müller Glia: Implications for retinal vulnerability

doi: 10.1016/j.redox.2025.103691

Figure Lengend Snippet: Knockdown of MT1G , AKAP12 and MAFF in human primary Müller glia. A. MT1G , AKAP12 and MAFF siRNA knockdown in primary Müller glia exposed to light stress. B & C . AlamarBlue cell viability assay on primary Müller glia with or without MT1G , AKAP12 and MAFF siRNA treatment in response to light stress. n = 6 biological replicates per group. Statistical analysis was performed using Welch's t -test (two-sided) between the control group (used consistently across all pairwise comparisons) and the respective knockdown groups. Data are presented as means ± standard error of the mean (SEM). D & E . JC1 assay on primary Müller glia with or without MT1G , AKAP12 and MAFF siRNA knockdown in response to light stress. n = 8 biological replicates per group. Statistical analysis was performed using Welch's t -test (two-sided) between the control group (used consistently across all pairwise comparisons) and the respective knockdown groups. F – H . Volcano plots of differential gene expression (p < 0.05, FC > 1.5) in MT1G , AKAP12 and MAFF siRNA knockdown vs. control groups. I – K . IPA of differential gene expression in human primary Müller glia with MT1G , AKAP12 and MAFF siRNA knockdown compared to the control group in response to light stress.

Article Snippet: Human primary Müller glia (P3) at 80 % confluency were transfected with MT1G (assay ID: s194623, ThermoFisher Scientific), AKAP12 (assay ID: s18435, ThermoFisher Scientific), MAFF (assay ID: s24370, ThermoFisher Scientific) and control small interfering RNA (siRNA) (assay ID: 4390843, ThermoFisher Scientific) at a concentration of 10 nM using Lipofectamine 3000 (#L3000-008, Invitrogen), following the manufacturer's instructions.

Techniques: Knockdown, Viability Assay, Control, Gene Expression