Journal: Cells

Article Title: HDAC1 -Mediated lncRNA Stimulatory Factor of Follicular Development to Inhibit the Apoptosis of Granulosa Cells and Regulate Sexual Maturity through miR-202-3p- COX1 Axis

doi: 10.3390/cells12232734

Figure Lengend Snippet: HDAC1 regulates the proliferation and apoptosis of GCs. ( A ) Effects TSA treatment at concentration of 0.01, 0.1, 0.25, 0.5, and 1 μM for 24 and 48 h on the viability of porcine GCs. ( B ) Micrographs of GCs treated with 0.1 and 0.25 μM TSA at 24 and 48 h. ( C ) Effects of 0.01, 0.1, 0.25, 0.5, and 1 μM TSA treatment on the expression of HDAC1 for 24 and 48 h. ( D – F ) The mRNA and protein levels of cell cycle and cell apoptosis-related genes in GCs treated with TSA. ( G , H ) The proliferation and apoptosis of GCs treated with TSA were assessed using EdU and flow cytometry. ( I ) Effect of TSA on the activity of Caspase 3/7 in porcine GCs was detected using Caspase 3/7 activity assay. ( J ) The interference efficiency of HDAC1 . ( K – M ) The mRNA and protein levels of cell cycle and cell apoptosis-related genes in GCs treated with HDAC1 -siRNA. ( N , O ) The proliferation and apoptosis of GCs treated with HDAC1 -siRNA were assessed using EdU and flow cytometry. ( P ) Effect of HDAC1 -siRNA on the activity of Caspase 3/7 in porcine GCs was detected using Caspase 3/7 activity assay. * indicates p < 0.05, ** indicates p < 0.01.

Article Snippet: The Caspase 3/7 Activity Assay was also performed to detect the apoptosis of GCs using the Caspase 3/7 Activity Assay Kit (Elabscience, Wuhan, China).

Techniques: Concentration Assay, Expressing, Flow Cytometry, Activity Assay

Journal: Cells

Article Title: HDAC1 -Mediated lncRNA Stimulatory Factor of Follicular Development to Inhibit the Apoptosis of Granulosa Cells and Regulate Sexual Maturity through miR-202-3p- COX1 Axis

doi: 10.3390/cells12232734

Figure Lengend Snippet: SFFD as ceRNA for miR-202-3p- COX1 regulates the proliferation and apoptosis of porcine GCs. ( A – D ) Effects of SFFD overexpression and miR-202-3p mimic on the mRNA and protein levels of genes related to cell cycle, E2 secretion, and apoptosis. ( E – H ) Effects of SFFD knockdown and miR-202-3p inhibitor on the mRNA and protein levels of genes related to cell cycle, E2 secretion, and apoptosis. ( I ) Effects of SFFD overexpression and miR-202-3p mimic on the proliferation of GCs. ( J ) Effects of SFFD knockdown and miR-202-3p inhibitor on the proliferation of GCs. ( K ) Effects of SFFD overexpression with miR-202-3p mimic and SFFD knockdown with miR-202-3p inhibitor on the E2 secretion of GCs. ( L , M ) Effects of SFFD overexpression and miR-202-3p mimic on the apoptosis and Caspase 3/7 activity of GCs. ( N , O ) Effects of SFFD knockdown and miR-202-3p inhibitor on the apoptosis and Caspase 3/7 activity of GCs. * indicates p < 0.05, ** indicates p < 0.01.

Article Snippet: The Caspase 3/7 Activity Assay was also performed to detect the apoptosis of GCs using the Caspase 3/7 Activity Assay Kit (Elabscience, Wuhan, China).

Techniques: Over Expression, Knockdown, Activity Assay

Journal: Experimental Animals

Article Title: Acyl-CoA thioesterase 1 (ACOT1) overexpression alleviates heart failure by inhibiting oxidative stress and cardiomyocyte apoptosis through the Kelch-like ECH-associated protein1-NF-E2-related factor2 (KEAP1-NRF2) pathway

doi: 10.1538/expanim.24-0129

Figure Lengend Snippet: (A) The levels of MDA and GSH and the activity of SOD in myocardial tissues were analyzed by ELISA. (B) The protein levels and the quantification results of cleaved PARP, cleaved CASPASE-3 and cleaved CASPASE-9 in myocardial tissues. (C) ROS in myocardial tissues were detected by DHE staining (n=6). The fluorescence intensity was calculated to evaluate ROS levels. bar=50 µ m. # P <0.05, ## P <0.01, ### P <0.001. n=6. EP: empty plasmid; ACOT1 oe : ACOT1 overexpression.

Article Snippet: The following antibodies were used: cleaved CASPASE-3 (Affinity, Cincinnati, OH, USA, #AF7022) diluted in 1:1,000; cleaved CASPASE-9 (Affinity, #AF5240) diluted in 1:1,000; cleaved PARP (ABclonal, Wuhan, China, #A22535) diluted in 1:1,000; ACOT1 (Novus Biologicals, Littleton, CO, USA, #NBP3-10843) diluted in 1:500; KEAP1 (Affinity, #AF5266) diluted in 1:1,000; NRF2 (Affinity, #AF0639) diluted in 1:1,000; Histone H3 (Gene Tex, Alton Parkway Irvine, CA, USA, #GTX122148) diluted in 1:5,000; GAPDH (Proteintech, Wuhan, China, #60004-1-Ig) diluted in 1:10,000; Goat Anti-Rabbit IgG/HRP (Solarbio, #SE134) diluted in 1:3,000; Goat Anti-Mouse IgG/HRP (Solarbio, #SE131) diluted in 1:3,000.

Techniques: Activity Assay, Enzyme-linked Immunosorbent Assay, Staining, Fluorescence, Plasmid Preparation, Over Expression

Journal: Experimental Animals

Article Title: Acyl-CoA thioesterase 1 (ACOT1) overexpression alleviates heart failure by inhibiting oxidative stress and cardiomyocyte apoptosis through the Kelch-like ECH-associated protein1-NF-E2-related factor2 (KEAP1-NRF2) pathway

doi: 10.1538/expanim.24-0129

Figure Lengend Snippet: (A) Protein level and the quantification of ACOT1 in HL-1 cells with ACOT1 overexpression. (B) cell viability of HL-1 cells with hypoxia treatment. (C) The levels of MDA and GSH and the activity of SOD in cells. (D) The protein levels of cleaved PARP, cleaved CASPASE-3 and cleaved CASPASE-9 in cells and the quantification results. (E) TUNEL staining of HL-1 cells and the positive cell quantification, bar=100 µ m. (F) ROS levels in HL-1 cells were detected by flow cytometry, MFI: mean fluorescence intensity. ## P <0.01, ### P <0.001. n=3. EP: empty plasmid; ACOT1 oe : ACOT1 overexpression.

Article Snippet: The following antibodies were used: cleaved CASPASE-3 (Affinity, Cincinnati, OH, USA, #AF7022) diluted in 1:1,000; cleaved CASPASE-9 (Affinity, #AF5240) diluted in 1:1,000; cleaved PARP (ABclonal, Wuhan, China, #A22535) diluted in 1:1,000; ACOT1 (Novus Biologicals, Littleton, CO, USA, #NBP3-10843) diluted in 1:500; KEAP1 (Affinity, #AF5266) diluted in 1:1,000; NRF2 (Affinity, #AF0639) diluted in 1:1,000; Histone H3 (Gene Tex, Alton Parkway Irvine, CA, USA, #GTX122148) diluted in 1:5,000; GAPDH (Proteintech, Wuhan, China, #60004-1-Ig) diluted in 1:10,000; Goat Anti-Rabbit IgG/HRP (Solarbio, #SE134) diluted in 1:3,000; Goat Anti-Mouse IgG/HRP (Solarbio, #SE131) diluted in 1:3,000.

Techniques: Over Expression, Activity Assay, TUNEL Assay, Staining, Flow Cytometry, Fluorescence, Plasmid Preparation

Journal: Translational Stroke Research

Article Title: Remote Ischemic Preconditioning Exerts Neuroprotective Effects Via the PGC-1α/FNDC5/BDNF Pathway in Focal Brain Ischemia of Rats

doi: 10.1007/s12975-026-01422-z

Figure Lengend Snippet: Effects of RIpreC on the expression of Bcl-2 ( A , B , F , G ), Bax ( A , C , F , H ), caspase-3 ( A , D , F , J ), and TUNEL-positive neurons ( A , E ) in the ischemic brain. Bcl-2 immunoreactivity in the perilesional cortex and its protein level were significantly increased in the RIpreC group compared the IR group. By contrast, Bax immunoreactivity and its protein level were significantly reduced in the RIpreC group compared with the IR group. The Bax/Bcl-2 ratios were also significantly reduced in the RIpreC group compared with the IR group ( I ). Subsequently, the number of TUNEL-positive neurons (arrow) and the ratios of caspase-3 positive to NeuN-positive neuron in the perilesional cortex were significantly decreased in the RIpreC group compared with the IR group ( E , K , L ). Mean ± standard error. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. Scale bar = 50 μm ( A ) and 25 μm ( K ). ( n = 4–11 in each group)

Article Snippet: # p < 0.05 (Mann–Whitney U test), # p < 0.01 (Student’s t-test), Scale bar = 1 mm ( B ). ( n = 5–11 in each group) Co-localization of mouse anti-neuronal nuclei (NeuN) antibody (1:200; a marker of neurons; ab104224; Abcam plc, Cambridge, UK) with rabbit anti-caspase-3 antibody (1:200; 19677-1-AP; Proteintech, USA) immunoreactivities was examined using immunofluorescence staining.

Techniques: Expressing, TUNEL Assay

Journal: Translational Stroke Research

Article Title: Remote Ischemic Preconditioning Exerts Neuroprotective Effects Via the PGC-1α/FNDC5/BDNF Pathway in Focal Brain Ischemia of Rats

doi: 10.1007/s12975-026-01422-z

Figure Lengend Snippet: Effects of PGC-1αInhibition by SR-18,292. Inhibition of PGC-1α blunted the neuroprotective and neurorestorative effects of RIpreC ( A – F ). No changes in brain infarctions, neurological scores, and sensorimotor functions were observed in the RIpreC-treated SR-18,292 group compared with the IR group. Immunohistochemical analysis of the rectangular areas in A showed that immunoreactivities of PGC-1α, FNDC5, BDNF, and caspase-3 surrounding the lesion in the RIpreC-treated SR-18,292 group were similar to those in the IR group ( G – K ). Scale bar = 50 μm ( G ). ( n = 5–11 in each group)

Article Snippet: # p < 0.05 (Mann–Whitney U test), # p < 0.01 (Student’s t-test), Scale bar = 1 mm ( B ). ( n = 5–11 in each group) Co-localization of mouse anti-neuronal nuclei (NeuN) antibody (1:200; a marker of neurons; ab104224; Abcam plc, Cambridge, UK) with rabbit anti-caspase-3 antibody (1:200; 19677-1-AP; Proteintech, USA) immunoreactivities was examined using immunofluorescence staining.

Techniques: Inhibition, Immunohistochemical staining

Journal: Frontiers in Immunology

Article Title: Tick lipocalin triggers mammalian IGFBP-3-mediated apoptosis in macrophages and keratinocytes

doi: 10.3389/fimmu.2026.1768484

Figure Lengend Snippet: rGST-Otlip treatment causes significant upregulation of apoptotic gene expression in murine raw macrophages. qRT-PCR analysis showing expression of igfbp-3R (A) , bax (B) , caspase-9 (C) , caspase-3 (D) , and bcl- 2 (E) in murine macrophage cells upon treatment with rGST or rGST-Otlip. The mRNA levels of cytokines were normalized to mouse actin mRNA levels. Statistical analysis was performed using Mann-Whitney U test and P value is shown. Open circles indicate data from samples generated upon treatment with rGST and closed circles represent data from samples generated upon treatment with rGST-Otlip. (F) Immunoblotting analysis showing levels of IGFBP3 and Caspase-3 in murine macrophage cells upon treatment with rGST or rGST-Otlip. Beta-actin levels serve as loading control in the immunoblotting analysis. Protein sizes are indicated as kilodaltons (kDa).

Article Snippet: After gel electrophoresis, blots were blocked with 5% milk buffer and probed with IGFBP3 rabbit polyclonal antibody (cat. no. A16052, ABclonal, USA) or Caspase-3 p12 Rabbit monoclonal antibody (cat. no. A19664, ABclonal, USA) or Bcl2 Rabbit polyclonal antibody (cat. vo.

Techniques: Gene Expression, Quantitative RT-PCR, Expressing, MANN-WHITNEY, Generated, Western Blot, Control

Journal: Frontiers in Immunology

Article Title: Tick lipocalin triggers mammalian IGFBP-3-mediated apoptosis in macrophages and keratinocytes

doi: 10.3389/fimmu.2026.1768484

Figure Lengend Snippet: Treatment of murine macrophages with O. turicata americanus salivary gland lysate induces apoptotic gene expression. qRT-PCR analysis showing expression of igfbp-3R (A) , bax (B) , caspase-9 (C) , caspase-3 (D) , and bcl-2 (E) transcripts in murine macrophages upon treatment with salivary gland lysates generated from O. turicata americanus unfed or fed nymphal ticks. The transcript levels of cytokines were normalized to mouse beta-actin transcript levels. Statistical analysis was performed using Mann-Whitney U test and P values are shown. Open circles indicate data from samples generated upon treatment with salivary gland lysates from unfed ticks and closed circles represent data from samples generated upon treatment with salivary gland lysates from fed ticks. (F) Immunoblotting analysis showing expression of IGFBP3 and Caspase-3 protein in murine macrophages upon treatment with O. turicata americanus unfed or fed salivary gland lysates. Beta-actin levels serve as loading control. Protein sizes are indicated as kilodaltons (kDa).

Article Snippet: After gel electrophoresis, blots were blocked with 5% milk buffer and probed with IGFBP3 rabbit polyclonal antibody (cat. no. A16052, ABclonal, USA) or Caspase-3 p12 Rabbit monoclonal antibody (cat. no. A19664, ABclonal, USA) or Bcl2 Rabbit polyclonal antibody (cat. vo.

Techniques: Gene Expression, Quantitative RT-PCR, Expressing, Generated, MANN-WHITNEY, Western Blot, Control

Journal: Frontiers in Immunology

Article Title: Tick lipocalin triggers mammalian IGFBP-3-mediated apoptosis in macrophages and keratinocytes

doi: 10.3389/fimmu.2026.1768484

Figure Lengend Snippet: Otlip treatment modulates apoptotic gene and protein expression in human keratinocytes: qRT-PCR analysis showing expression of igfbp-3 (A) , caspase-3 (B) and bcl-2 (C) in HaCaT cells upon treatment with rGST or rGST-Otlip. The mRNA levels of cytokines were normalized to human beta-actin mRNA levels. Open circles indicate data from samples generated upon treatment with rGST and closed circles represent data from samples generated upon treatment with rGST-Otlip. Statistical analysis was performed using Mann-Whitney U test and P value is shown. (D) Immunoblotting analysis showing expression of IGFBP3, Caspase -3 and BCL-2 proteins in HaCaT cells upon treatment with rGST or rGST-Otlip. Beta-actin levels serve as loading controls. Protein sizes are indicated as kilodaltons (kDa).

Article Snippet: After gel electrophoresis, blots were blocked with 5% milk buffer and probed with IGFBP3 rabbit polyclonal antibody (cat. no. A16052, ABclonal, USA) or Caspase-3 p12 Rabbit monoclonal antibody (cat. no. A19664, ABclonal, USA) or Bcl2 Rabbit polyclonal antibody (cat. vo.

Techniques: Expressing, Quantitative RT-PCR, Generated, MANN-WHITNEY, Western Blot

Journal: Frontiers in Immunology

Article Title: Tick lipocalin triggers mammalian IGFBP-3-mediated apoptosis in macrophages and keratinocytes

doi: 10.3389/fimmu.2026.1768484

Figure Lengend Snippet: Knockdown of igfbp3 expression affects rGST-Otlip-induced apoptosis in murine macrophages. qRT-PCR analysis showing expression of igfbp3 (A) , caspase-3 (B) and bcl-2 (C) in igfbp3/scrambled -siRNA-rGST-Otlip-treated macrophages. The mRNA levels of igfbp3 (A) , caspase-3 (B) and bcl-2 (C) were normalized to mouse beta-actin mRNA levels. Open circles indicate data from scrambled siRNA-treated group and closed circles represent data from igfbp3 -siRNA-treated group. Statistical analysis was performed using Mann-Whitney U test and P values are shown. (D) Immunoblotting analysis showing expression of IGFBP3 and Caspase -3 proteins in igfbp3/scrambled -siRNA-rGST-Otlip-treated macrophages. Mouse beta-actin levels serve as loading controls. Protein sizes are indicated as kilodaltons (kDa). (E) Schematic representation showing the role of O. turicata americanus lipocalin in the modulation of apoptosis via IGFBP3 signaling in mammalian cells. While feeding on a mammalian host, ticks release salivary cocktail containing histamine-binding lipocalin. The secreted tick lipocalin stimulates the production of IGFBP-3 (Insulin-like growth factor-binding protein-3) and its receptor IGFBP-3R in both murine macrophages and human keratinocytes. The secreted IGFBP-3 could then bind to IGFBP-3R to stimulate apoptotic cascade. Otlip induces mammalian Caspase 9 (initiator caspase), Caspase 3 (executioner caspase) and repress BCL-2 (anti-apoptotic) to induce apoptosis in macrophages and keratinocytes. Induction in apoptosis of immune cells like macrophages and resident cells like keratinocytes would enable ticks to successfully complete their blood feeding.

Article Snippet: After gel electrophoresis, blots were blocked with 5% milk buffer and probed with IGFBP3 rabbit polyclonal antibody (cat. no. A16052, ABclonal, USA) or Caspase-3 p12 Rabbit monoclonal antibody (cat. no. A19664, ABclonal, USA) or Bcl2 Rabbit polyclonal antibody (cat. vo.

Techniques: Knockdown, Expressing, Quantitative RT-PCR, MANN-WHITNEY, Western Blot, Binding Assay