sj-pdf-3-tva-10.1177_15248380231151690 – Supplemental material for The Effectiveness of Parenting Programs in Preventing Abusive Head Trauma: A Systematic Review and Meta-Analysis

Supplemental material, sj-pdf-3-tva-10.1177_15248380231151690 for The Effectiveness of Parenting Programs in Preventing Abusive Head Trauma: A Systematic Review and Meta-Analysis by Hsin-Yi Chang, Yu-Chun Chang, Yi-Ting Chang, Yi-Wen Chen, Pei-Yu Wu and Jui-Ying Feng in Trauma, Violence, & Abuse</p

sj-pdf-1-tva-10.1177_15248380231151690 – Supplemental material for The Effectiveness of Parenting Programs in Preventing Abusive Head Trauma: A Systematic Review and Meta-Analysis

Supplemental material, sj-pdf-1-tva-10.1177_15248380231151690 for The Effectiveness of Parenting Programs in Preventing Abusive Head Trauma: A Systematic Review and Meta-Analysis by Hsin-Yi Chang, Yu-Chun Chang, Yi-Ting Chang, Yi-Wen Chen, Pei-Yu Wu and Jui-Ying Feng in Trauma, Violence, & Abuse</p

sj-pdf-2-tva-10.1177_15248380231151690 – Supplemental material for The Effectiveness of Parenting Programs in Preventing Abusive Head Trauma: A Systematic Review and Meta-Analysis

Supplemental material, sj-pdf-2-tva-10.1177_15248380231151690 for The Effectiveness of Parenting Programs in Preventing Abusive Head Trauma: A Systematic Review and Meta-Analysis by Hsin-Yi Chang, Yu-Chun Chang, Yi-Ting Chang, Yi-Wen Chen, Pei-Yu Wu and Jui-Ying Feng in Trauma, Violence, & Abuse</p

Schematic illustration of chemical structures and docking stimulation outlining.

<p>(A, C) c(RGDyK) and (B,D) E[c(RGDyK)]₂ peptides, respectively.</p

Pathways of caspase cascade and cytochrome c release are involved in U87MG cells treated with RGD peptides in the presence or absence of 10 nM PTX.

<p>The low-dose PTX induced the activation of mitotic arrest that is the main cellular mechanism of programmed cell death, and subsequently increased the expression of caspases -10, -12 and -8 for apoptosis (black dashed line). The binding of RGD peptides to integrin-αvβ3 activates caspase -9 then activates downstream caspase -8, -10 and executor caspase -3 (black bold line). Integrin-αvβ3 attached with normal ECM and directed cells to growth or proliferation (the bold gray line). Numbers 1 and 3 represented U87MG cells pre-treated by low-dose PTX or treated by RGD peptides, respectively. Number 2 indicates the recruited phenomenon of integrin-αvβ3 after low-dose PTX pre-treatment. MT: microtubules; ECM: extracellular cell matrix.</p

Expression of surface integrin-αvβ3 in U87MG cells with the treatment of 10 nM PTX.

<p>(A) Cytometric analysis of the expression of integrin-αvβ3 was determined by specific integrin-αvβ3 antibody staining. The expression of integrin-αvβ3 in U87MG cells treated with 10 nM PTX for 12 hrs was higher than control group. (B) Immunofluorescent detection of integrin-αvβ3 and nuclei was respectively stained by anti-integrin-αvβ3 antibody (Texas red) and DAPI (blue). Integrin-αvβ3 was obviously localized in the cell periphery. (I-III) indicated the U87MG cell isotypic control group. (IV-VI) Integrin-αvβ3 of U87MG cells was induced after 12 hr 10 nM PTX. (VII–IX) U87MG cells were pre-incubated with E[c(RGDyK)]₂ peptide to antagonized PTX-induced integrin-αvβ3 expression.</p

Primers used in real-time PCR analysis.

<p>Primers used in real-time PCR analysis.</p

Cytotoxicity of U87MG cells treated with RGD peptide or the combination of PTX and RGD peptide was quantified using MTT assay.

<p>(A) Cell viability of c(RGDyK) peptide or combination pre-treated by 10 nM PTX in U87MG cells. (B) Cell viability of E[c(RGDyK)]₂ peptides or combination pre-treated by 10 nM PTX in U87MG cells. Cell viability was decreased at 100 µM c(RGDyK) or E[c(RGDyK)]₂ peptide. (C) Cell viability of a serial concentration of PTX for 24 hr in U87MG cells. The results of triplicate experiments are expressed as mean ± standard deviation.</p

Cytometric analysis of apoptosis events in U87MG cells treated with PTX, c(RGDyK)/E[c(RGDyK)]₂ peptides or the combination of PTX and c(RGDyK)/E[c(RGDyK)]₂ peptides.

<p>Determination of apoptosis was using Annexin-V-FITC staining and FACS detector. The occurrence of programmed apoptosis was obviously in U87MG cells pre-treated with 10 nM PTX followed by 100 µM (A) c(RGDyK)/(B) E[c(RGDyK)]₂ peptides. Triplicate experiments were obtained for analysis.</p

Neutralized effect of caspase-3, -8 and -9 inhibitors on the caspase mRNA expression induced by PTX pre-treatment and E[c(RGDyK)]₂ peptide.

<p>U87MG cells were respectively pre-incubated with inhibitors of caspase-3 (Z-DEVD-FMK), caspase-8 (Z-IETD-FMK) or caspase-9 (Z-LEHD-FMK) and followed by 10 nM PTX pre-treatment for 12 hr and 40 µM E[c(RGDyK)]₂ treatment for 24 hr. The mRNA expression of (A) caspase-3, (B) caspase-8, (C) caspase-9, (D) caspase-10, and (E) caspase-12 was determined using real-time PCR. Eventually, either caspase-3, -8 or -9 inhibitor effectively to suppress the expression of caspase-3, -8 and -9. Asterisk indicates significant at p<0.05 compared with 10 nM PTX alone.</p