Journal: iScience

Article Title: Human OPN-derived peptide SV prevents BPD model through interacting with KIF5B to repair mitochondrial damage and inhibit apoptosis

doi: 10.1016/j.isci.2025.113691

Figure Lengend Snippet: SV repairs LPS-induced cell damage in vitro may rely on the inhibition of apoptosis (A) The CCK8 assay detected the proliferation of cells in the control, LPS (100 ng/mL), and LPS+SV (100 μM) groups, N = 3. (B) Flow assay of ROS levels in MLE-12 cells from control, LPS, and LPS+SV groups, N = 3. (C) Apoptosis of MLE-12 cells in the control, LPS, and LPS+SV groups was detected using flow cytometry, N = 3. (D) qPCR analysis showed the mRNA levels of Bcl-2, Bax, SPC, IL-1β, and IL-6, N = 6. (E and F) Western blot revealed the protein expression of Bcl-2, Bax, SPC, and VEGFA (vascular endothelial growth factor) in MLE-12 cells of in three groups, N = 3. Unpaired t test, ∗/#/ p < 0.05,∗∗/##/ p < 0.01, and ∗∗∗/ p < 0.001. CTL, control; LPS, BPD induced by lipopolysaccharide.

Article Snippet: Apoptosis was evaluated using an Annexin V-FITC/PI Apoptosis Detection Kit (Vazyme, China).

Techniques: In Vitro, Inhibition, CCK-8 Assay, Control, Flow Cytometry, Western Blot, Expressing

Journal: iScience

Article Title: Human OPN-derived peptide SV prevents BPD model through interacting with KIF5B to repair mitochondrial damage and inhibit apoptosis

doi: 10.1016/j.isci.2025.113691

Figure Lengend Snippet: SV repairs hyperoxia-induced cell damage in vitro may rely on the inhibition of apoptosis (A) The CCK8 assay detected the proliferation of cells in the control, hyperoxia (Hyp), and hyperoxia with SV (100 μM) groups, N = 3. (B) Flow assay of ROS levels in MLE-12 cells from control, Hyp, and Hyp+SV groups, N = 3. (C) Apoptosis of MLE-12 cells in the control, Hyp, and Hyp+SV groups was detected using flow cytometry, N = 3. (D) mRNA expression of Bcl-2, Bax, SPC, IL-1β, and IL-6 in three groups of MLE-12 cells, N = 6. (E and F) Western blot revealed the protein expression of Bcl-2, Bax, SPC, and VEGFA in MLE-12 cells of in three groups, N = 3. Unpaired t test, ∗/#/ p < 0.05,∗∗/##/ p < 0.01, and ∗∗∗/###/ p < 0.001. CTL, control; Hyp, BPD induced by hyperoxia.

Article Snippet: Apoptosis was evaluated using an Annexin V-FITC/PI Apoptosis Detection Kit (Vazyme, China).

Techniques: In Vitro, Inhibition, CCK-8 Assay, Control, Flow Cytometry, Expressing, Western Blot

Journal: iScience

Article Title: Human OPN-derived peptide SV prevents BPD model through interacting with KIF5B to repair mitochondrial damage and inhibit apoptosis

doi: 10.1016/j.isci.2025.113691

Figure Lengend Snippet: SV improves the LPS-induced BPD animal model (A) Body weight changes from 1 to 7 days in control, LPS, and LPS+SV groups of mice, N = 6. (B) Lung morphology of control, LPS, and LPS+SV groups, N = 6. (C) Lung histopathologic changes (H&E), N = 6. Mean linear intercept (MLI) quantitative represents the mean airspace diameter and mean alveolar number (MAN) quantitative represents the mean number of alveoli. Scale bars: 500 μm/100 μm. (D) Representative images of TUNEL immunostaining showing the apoptosis (green staining) in rat lung tissue of control, LPS and LPS+SV groups, N = 6. Representative images of Ki67 immunostaining showing the proliferation (red staining) in rat lung tissue of control, LPS, and LPS+SV groups; N = 6. Scale bars: 500 μm. (E) qPCR analysis showed the mRNA levels of Bcl-2, Bax, SPC, IL-1β, and IL-6, N = 6. (F and G) Western blot showed the protein levels of Bcl-2, Bax, SPC, and VEGFA, N = 6. Unpaired t test, ∗/#/ p < 0.05,∗∗/##/ p < 0.01, and ∗∗∗/###/ p < 0.001. CTL, control; LPS, BPD induced by lipopolysaccharide. The LPS concentration was 500 μg/kg and the SV intervention concentration was 200 μg/kg depending on the weight of the pups per day.

Article Snippet: Apoptosis was evaluated using an Annexin V-FITC/PI Apoptosis Detection Kit (Vazyme, China).

Techniques: Animal Model, Control, TUNEL Assay, Immunostaining, Staining, Western Blot, Concentration Assay

Journal: iScience

Article Title: Human OPN-derived peptide SV prevents BPD model through interacting with KIF5B to repair mitochondrial damage and inhibit apoptosis

doi: 10.1016/j.isci.2025.113691

Figure Lengend Snippet: SV improves the hyperoxia-induced BPD animal model (A) Body weight changes from 1 to 7 days in control, Hyp, and Hyp+SV groups of mice, N = 6. (B) Lung morphology of control, Hyp, and Hyp+SV groups of mice, N = 6. (C) Lung histopathologic changes (H&E), N = 6. MLI quantitative represents the mean airspace diameter and MAN quantitative represents the mean number of alveoli. Scale bars: 500 μm/100 μm. (D) Representative images of TUNEL immunostaining showing the apoptosis (green staining) in rat lung tissue of control, Hyp, and Hyp+SV groups. Representative images of Ki67 immunostaining showing the proliferation (red staining) in rat lung tissue of control, LPS, and LPS+SV groups, N = 6. Scale bars: 500 μm. (E) qPCR analysis showed the mRNA levels of Bcl-2, Bax, SPC, IL-1β, and IL-6, N = 6. (F and G) Western blot showed the protein levels of Bcl-2, Bax, SPC, and VEGFA, N = 6. Unpaired t test, ∗/#/ p < 0.05,∗∗/##/ p < 0.01, and ∗∗∗/###/ p < 0.001. CTL, control; Hyp, BPD induced by hyperoxia. The SV intervention concentration was 200 μg/kg depending on the weight of the pups per day.

Article Snippet: Apoptosis was evaluated using an Annexin V-FITC/PI Apoptosis Detection Kit (Vazyme, China).

Techniques: Animal Model, Control, TUNEL Assay, Immunostaining, Staining, Western Blot, Concentration Assay

Journal: Journal of controlled release : official journal of the Controlled Release Society

Article Title: RNA-based micelles: A novel platform for paclitaxel loading and delivery

doi: 10.1016/j.jconrel.2018.02.014

Figure Lengend Snippet: A. Assay cytotoxicity effects of pRNA-3WJ-PTX micelles by MTT assay. B. Assay apoptotic effects of pRNA-3WJ-PTX micelles by PI/Annexin V-FITC dual staining and FACS analysis. C. Caspase-3 assay.

Article Snippet: Apoptosis study in in vitro cell model FITC Annexin V Apoptosis Detection Kit (BD Pharmingen) and Caspase-3 Assay Kit (BD Pharmingen) were used as previously reported [ 64 ] to study cell apoptosis induced by pRNA-3WJ-PTX micelles treatment.

Techniques: MTT Assay, Staining, Caspase-3 Assay

Journal: World Journal of Surgical Oncology

Article Title: Circ_0026416 downregulation blocks the development of colorectal cancer through depleting MYO6 expression by enriching miR-545-3p

doi: 10.1186/s12957-021-02407-y

Figure Lengend Snippet: Circ_0026416 downregulation inhibited CRC development in vitro and in vivo. A The expression of circ_0026416 in HCT-8 and SW480 cells after si-circ_0026416 transfection was checked by qPCR. B , C Cell proliferation was checked by CCK-8 assay. D Cell proliferation was also checked by colony formation assay. E , F Cell migration and cell invasion were examined using Transwell assay. G The ability of angiogenesis was checked by tube formation assay. H Cell apoptosis was examined using flow cytometry assay. I , J The protein levels of E-Cadherin, vimentin, and N-Cadherin were determined by western blot. K – M Animal study was performed to determine the role of circ_0026416 knockdown in vivo. N The expression of circ_0026416 in tumor tissues from animal model was checked by qPCR. * P < 0.05

Article Snippet: At 48 h post-transfection, the Annexin V-FITC Apoptosis Detection Kit (Beyotime, Shanghai, China) was used to monitor cell apoptosis using a flow cytometer (BD Biosciences) according to the protocol from kit.

Techniques: In Vitro, In Vivo, Expressing, Transfection, CCK-8 Assay, Colony Assay, Migration, Transwell Assay, Tube Formation Assay, Flow Cytometry, Western Blot, Knockdown, Animal Model

Journal: World Journal of Surgical Oncology

Article Title: Circ_0026416 downregulation blocks the development of colorectal cancer through depleting MYO6 expression by enriching miR-545-3p

doi: 10.1186/s12957-021-02407-y

Figure Lengend Snippet: Circ_0026416 knockdown inhibited CRC cell development by enriching miR-545-3p. In function rescue experiments, A , B cell proliferation was assessed by CCK-8 assay. C Colony formation assay was performed to monitor cell proliferation. D , E Cell migration and cell invasion were determined by Transwell assay. F The ability of angiogenesis was checked by tube formation assay. G Flow cytometry assay was conducted to assess cell apoptosis. H , I The protein levels of E-Cadherin, vimentin, and N-Cadherin were quantified by western blot. * P < 0.05

Article Snippet: At 48 h post-transfection, the Annexin V-FITC Apoptosis Detection Kit (Beyotime, Shanghai, China) was used to monitor cell apoptosis using a flow cytometer (BD Biosciences) according to the protocol from kit.

Techniques: Knockdown, CCK-8 Assay, Colony Assay, Migration, Transwell Assay, Tube Formation Assay, Flow Cytometry, Western Blot

Journal: World Journal of Surgical Oncology

Article Title: Circ_0026416 downregulation blocks the development of colorectal cancer through depleting MYO6 expression by enriching miR-545-3p

doi: 10.1186/s12957-021-02407-y

Figure Lengend Snippet: MiR-545-3p restoration blocked CRC cell malignant behaviors by targeting MYO6. In function rescue experiments, A , B cell proliferation was assessed by CCK-8 assay. C Cell proliferation was assessed by colony formation assay. D , E Cell migration and cell invasion were examined using Transwell assay. F The ability of angiogenesis was checked by tube formation assay. G Flow cytometry assay was conducted to assess cell apoptosis. H , I The protein levels of E-Cadherin, vimentin and N-Cadherin were quantified by western blot. * P < 0.05

Article Snippet: At 48 h post-transfection, the Annexin V-FITC Apoptosis Detection Kit (Beyotime, Shanghai, China) was used to monitor cell apoptosis using a flow cytometer (BD Biosciences) according to the protocol from kit.

Techniques: CCK-8 Assay, Colony Assay, Migration, Transwell Assay, Tube Formation Assay, Flow Cytometry, Western Blot

Journal: Molecular Cancer Therapeutics

Article Title: Inflexinol inhibits colon cancer cell growth through inhibition of nuclear factor-κB activity via direct interaction with p50

doi: 10.1158/1535-7163.mct-08-0694

Figure Lengend Snippet: Figure 3. Apoptotic cell death and expression of apoptosis-related proteins of colon cancer cells by inflexinol. A, colon cancer cells were treated with several concentration of inflexinol for 24 h; cell morphologic changes were observed under a microscope (top); and apoptotic cells were examined with a fluorescence microscope after TUNEL staining (bottom). Total number of cells in a given area was determined by DAPI nuclear staining (fluorescent mi- croscope; middle). The apoptotic index was determined as the number of DAPI-stained TUNEL-positive cells counted. Columns, mean of three experiments each done in triplicate; bars, SD. *, P < 0.05, versus the untreated group. B, the cells were treated with different concentrations (10–40 μmol/L) of inflexinol at 37°C for 24 h. Equal amounts of total proteins (50 μg/lane) were subjected to 12% SDS-PAGE. Expression of Bax, cleaved caspase-3, cleaved caspase-9, cleaved poly(ADP-ribose) polymerase (PARP), cyclin D1 (CyD1), Bcl-2, XIAP, cIAP1/2, and β-actin were detected by Western blotting using specific antibodies. β-Actin protein was used as an internal control. Each blot is representative of three independent experimental results. Bar, 100 μm.

Article Snippet: The membrane was incubated for 5 h at room temperature with specific antibodies: mouse polyclonal antibodies against p65 and p50 (1:500 dilution; Santa Cruz Biotechnology, Inc.) and Ki-67 (1:500 dilution; Dakocytomation) and rabbit polyclonal for Bax and Bcl-2 (1:500 dilution; Santa Cruz Biotechnology), caspase-3, cleaved caspase-3, cleaved caspase-9, poly(ADP-ribose) polymerase, antihuman proliferating cell nuclear antigen (PCNA), X-chromosome linked inhibitor of apoptosis protein (XIAP), and cellular inhibitor of apoptosis protein 1 (cIAP1; 1:1,000 dilution; Cell Signaling Technology, Inc.).

Techniques: Expressing, Concentration Assay, Microscopy, Fluorescence, TUNEL Assay, Staining, SDS Page, Western Blot, Control

Journal: Molecular Cancer Therapeutics

Article Title: Inflexinol inhibits colon cancer cell growth through inhibition of nuclear factor-κB activity via direct interaction with p50

doi: 10.1158/1535-7163.mct-08-0694

Figure Lengend Snippet: Figure 7. NF-κB activity and expression of NF-κB and apoptosis-related proteins as well as induction of apoptosis by inflexinol in in vivo xenograft animal model. A, DNA binding activity of NF-κB was determined by EMSA in nuclear extract from xenograft tumor samples (five samples per group) as described in Materials and Methods. B, expression of NF-κB proteins was detected by Western blotting using specific antibodies in nuclear extract from xenograft tumor samples (three samples per group) as described in Materials and Methods. C, immunohistochemistry was used to determine the nuclear p65 and p50 reactive cell number in nude mouse xenograft tissues by the different treatments as described in Materials and Methods. A representative sample from each group was stained in the picture. D, Western blot analysis of cleaved caspase-3, Bax, Bcl-2, IAP1/2, and XIAP expression. E, immunohistochemical analysis of cleaved caspase-3. F, apoptotic cell death by inflexinol in vivo. Columns, mean from five animal tumor sections; bars, SD. *, P < 0.05, versus the control group. Bar, 100 μm.

Article Snippet: The membrane was incubated for 5 h at room temperature with specific antibodies: mouse polyclonal antibodies against p65 and p50 (1:500 dilution; Santa Cruz Biotechnology, Inc.) and Ki-67 (1:500 dilution; Dakocytomation) and rabbit polyclonal for Bax and Bcl-2 (1:500 dilution; Santa Cruz Biotechnology), caspase-3, cleaved caspase-3, cleaved caspase-9, poly(ADP-ribose) polymerase, antihuman proliferating cell nuclear antigen (PCNA), X-chromosome linked inhibitor of apoptosis protein (XIAP), and cellular inhibitor of apoptosis protein 1 (cIAP1; 1:1,000 dilution; Cell Signaling Technology, Inc.).

Techniques: Activity Assay, Expressing, In Vivo, Animal Model, Binding Assay, Western Blot, Immunohistochemistry, Staining, Immunohistochemical staining, Control