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Effects of PAQR3 overexpression on ferroptosis in HCC cells. (a) <t>Lipid</t> <t>peroxidation</t> levels measured by <t>TBARS</t> assay. (b, c) ROS levels detected by C11-BODIPY staining (red/green ratio; scale bars: 50 μm). (d, e) Intracellular Fe 2+ levels assessed by fluorescence intensity (scale bars: 50 μm). (f) Representative western blot bands of ferroptosis-related proteins (SLC7A11, GPX4, ACSL4, and b-actin). (g) Quantification of relative SLC7A11 expression. (h) Quantification of relative GPX4 expression. (i) Quantification of relative ACSL4 expression. ✶ ✶ ✶ P < 0.001, ns: Not significant. Data in (a, g, h, i) are presented as mean±standard deviation from three independent experiments. Data in (b-e) and blot in (f) are representative of three independent experiments. PAQR3: Progestin and adipoQ receptor 3, HCC: Hepatocellular carcinoma, TBARS: Thiobarbituric acid reactive substance, ROS: Reactive oxygen species, Oe-NC: Overexpression negative control.
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Effects of PAQR3 overexpression on ferroptosis in HCC cells. (a) <t>Lipid</t> <t>peroxidation</t> levels measured by <t>TBARS</t> assay. (b, c) ROS levels detected by C11-BODIPY staining (red/green ratio; scale bars: 50 μm). (d, e) Intracellular Fe 2+ levels assessed by fluorescence intensity (scale bars: 50 μm). (f) Representative western blot bands of ferroptosis-related proteins (SLC7A11, GPX4, ACSL4, and b-actin). (g) Quantification of relative SLC7A11 expression. (h) Quantification of relative GPX4 expression. (i) Quantification of relative ACSL4 expression. ✶ ✶ ✶ P < 0.001, ns: Not significant. Data in (a, g, h, i) are presented as mean±standard deviation from three independent experiments. Data in (b-e) and blot in (f) are representative of three independent experiments. PAQR3: Progestin and adipoQ receptor 3, HCC: Hepatocellular carcinoma, TBARS: Thiobarbituric acid reactive substance, ROS: Reactive oxygen species, Oe-NC: Overexpression negative control.
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Effects of PAQR3 overexpression on ferroptosis in HCC cells. (a) <t>Lipid</t> <t>peroxidation</t> levels measured by <t>TBARS</t> assay. (b, c) ROS levels detected by C11-BODIPY staining (red/green ratio; scale bars: 50 μm). (d, e) Intracellular Fe 2+ levels assessed by fluorescence intensity (scale bars: 50 μm). (f) Representative western blot bands of ferroptosis-related proteins (SLC7A11, GPX4, ACSL4, and b-actin). (g) Quantification of relative SLC7A11 expression. (h) Quantification of relative GPX4 expression. (i) Quantification of relative ACSL4 expression. ✶ ✶ ✶ P < 0.001, ns: Not significant. Data in (a, g, h, i) are presented as mean±standard deviation from three independent experiments. Data in (b-e) and blot in (f) are representative of three independent experiments. PAQR3: Progestin and adipoQ receptor 3, HCC: Hepatocellular carcinoma, TBARS: Thiobarbituric acid reactive substance, ROS: Reactive oxygen species, Oe-NC: Overexpression negative control.
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Effects of PAQR3 overexpression on ferroptosis in HCC cells. (a) <t>Lipid</t> <t>peroxidation</t> levels measured by <t>TBARS</t> assay. (b, c) ROS levels detected by C11-BODIPY staining (red/green ratio; scale bars: 50 μm). (d, e) Intracellular Fe 2+ levels assessed by fluorescence intensity (scale bars: 50 μm). (f) Representative western blot bands of ferroptosis-related proteins (SLC7A11, GPX4, ACSL4, and b-actin). (g) Quantification of relative SLC7A11 expression. (h) Quantification of relative GPX4 expression. (i) Quantification of relative ACSL4 expression. ✶ ✶ ✶ P < 0.001, ns: Not significant. Data in (a, g, h, i) are presented as mean±standard deviation from three independent experiments. Data in (b-e) and blot in (f) are representative of three independent experiments. PAQR3: Progestin and adipoQ receptor 3, HCC: Hepatocellular carcinoma, TBARS: Thiobarbituric acid reactive substance, ROS: Reactive oxygen species, Oe-NC: Overexpression negative control.
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Effects of PAQR3 overexpression on ferroptosis in HCC cells. (a) <t>Lipid</t> <t>peroxidation</t> levels measured by <t>TBARS</t> assay. (b, c) ROS levels detected by C11-BODIPY staining (red/green ratio; scale bars: 50 μm). (d, e) Intracellular Fe 2+ levels assessed by fluorescence intensity (scale bars: 50 μm). (f) Representative western blot bands of ferroptosis-related proteins (SLC7A11, GPX4, ACSL4, and b-actin). (g) Quantification of relative SLC7A11 expression. (h) Quantification of relative GPX4 expression. (i) Quantification of relative ACSL4 expression. ✶ ✶ ✶ P < 0.001, ns: Not significant. Data in (a, g, h, i) are presented as mean±standard deviation from three independent experiments. Data in (b-e) and blot in (f) are representative of three independent experiments. PAQR3: Progestin and adipoQ receptor 3, HCC: Hepatocellular carcinoma, TBARS: Thiobarbituric acid reactive substance, ROS: Reactive oxygen species, Oe-NC: Overexpression negative control.
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Effects of PAQR3 overexpression on ferroptosis in HCC cells. (a) Lipid peroxidation levels measured by TBARS assay. (b, c) ROS levels detected by C11-BODIPY staining (red/green ratio; scale bars: 50 μm). (d, e) Intracellular Fe 2+ levels assessed by fluorescence intensity (scale bars: 50 μm). (f) Representative western blot bands of ferroptosis-related proteins (SLC7A11, GPX4, ACSL4, and b-actin). (g) Quantification of relative SLC7A11 expression. (h) Quantification of relative GPX4 expression. (i) Quantification of relative ACSL4 expression. ✶ ✶ ✶ P < 0.001, ns: Not significant. Data in (a, g, h, i) are presented as mean±standard deviation from three independent experiments. Data in (b-e) and blot in (f) are representative of three independent experiments. PAQR3: Progestin and adipoQ receptor 3, HCC: Hepatocellular carcinoma, TBARS: Thiobarbituric acid reactive substance, ROS: Reactive oxygen species, Oe-NC: Overexpression negative control.

Journal: CytoJournal

Article Title: A novel candidate tumor biomarker progestin and adipoQ receptor 3 regulates cell metastasis through transforming growth factor-β pathway in hepatocellular carcinoma

doi: 10.25259/Cytojournal_138_2025

Figure Lengend Snippet: Effects of PAQR3 overexpression on ferroptosis in HCC cells. (a) Lipid peroxidation levels measured by TBARS assay. (b, c) ROS levels detected by C11-BODIPY staining (red/green ratio; scale bars: 50 μm). (d, e) Intracellular Fe 2+ levels assessed by fluorescence intensity (scale bars: 50 μm). (f) Representative western blot bands of ferroptosis-related proteins (SLC7A11, GPX4, ACSL4, and b-actin). (g) Quantification of relative SLC7A11 expression. (h) Quantification of relative GPX4 expression. (i) Quantification of relative ACSL4 expression. ✶ ✶ ✶ P < 0.001, ns: Not significant. Data in (a, g, h, i) are presented as mean±standard deviation from three independent experiments. Data in (b-e) and blot in (f) are representative of three independent experiments. PAQR3: Progestin and adipoQ receptor 3, HCC: Hepatocellular carcinoma, TBARS: Thiobarbituric acid reactive substance, ROS: Reactive oxygen species, Oe-NC: Overexpression negative control.

Article Snippet: Lipid peroxidation in HCC cells following the indicated treatments was assessed using a TBARS Assay Kit (Elabscience Biotechnology Co., Ltd., E-BC-K298-M, Wuhan, Hubei, China).

Techniques: Over Expression, TBARS Assay, Staining, Fluorescence, Western Blot, Expressing, Standard Deviation, Negative Control

PAQR3 regulates HCC cell metastasis and ferroptosis through the TGF-b pathway. (a) Wound healing assay images showing cell migration in different groups (scale bars: 200 μm). (b) Quantification of cell migration rates from wound healing assay. (c) Transwell invasion assay images of HCC cells in different groups (scale bars: 50 μm). (d) Quantification of invaded cell counts from the Transwell assay. (e) Representative Western blot images of EMT-related proteins (E-cadherin, N-cadherin, Vimentin, b-actin). (f) Quantification of relative E-cadherin expression. (g) Quantification of relative N-cadherin expression. (h) Quantification of relative Vimentin expression. (i) TBARS assay showing lipid peroxidation levels. (j) C11-BODIPY staining showing ROS levels (merge, oxidized C11, non-oxidized C11, scale bars: 50 μm). (k) Representative images of intracellular Fe 2+ fluorescence (scale bars: 50 μm). (l) Quantification of red/green fluorescence ratio for ROS levels. (m) Quantification of intracellular Fe 2+ fluorescence intensity. (n) Representative western blot images of ferroptosis-related proteins (SLC7A11, GPX4, ACSL4, b-actin). (o) Quantification of relative SLC7A11 expression. (p) Quantification of relative GPX4 expression. (q) Quantification of relative ACSL4 expression. ✶ P < 0.05, ✶ ✶ P < 0.01, ✶ ✶ ✶ P < 0.001, ns: Not significant. Images in (a, c, e, j, k, n) are representative of three independent experiments. Quantitative data in (b, d, f-h, i, l, m, o-q) are presented as mean±SD from three independent experiments. PAQR3: Progestin and adipoQ receptor 3, HCC: Hepatocellular carcinoma, EMT: Epithelial-mesenchymal transition, TBARS: Thiobarbituric acid reactive substance, Oe-NC: Overexpression negative control.

Journal: CytoJournal

Article Title: A novel candidate tumor biomarker progestin and adipoQ receptor 3 regulates cell metastasis through transforming growth factor-β pathway in hepatocellular carcinoma

doi: 10.25259/Cytojournal_138_2025

Figure Lengend Snippet: PAQR3 regulates HCC cell metastasis and ferroptosis through the TGF-b pathway. (a) Wound healing assay images showing cell migration in different groups (scale bars: 200 μm). (b) Quantification of cell migration rates from wound healing assay. (c) Transwell invasion assay images of HCC cells in different groups (scale bars: 50 μm). (d) Quantification of invaded cell counts from the Transwell assay. (e) Representative Western blot images of EMT-related proteins (E-cadherin, N-cadherin, Vimentin, b-actin). (f) Quantification of relative E-cadherin expression. (g) Quantification of relative N-cadherin expression. (h) Quantification of relative Vimentin expression. (i) TBARS assay showing lipid peroxidation levels. (j) C11-BODIPY staining showing ROS levels (merge, oxidized C11, non-oxidized C11, scale bars: 50 μm). (k) Representative images of intracellular Fe 2+ fluorescence (scale bars: 50 μm). (l) Quantification of red/green fluorescence ratio for ROS levels. (m) Quantification of intracellular Fe 2+ fluorescence intensity. (n) Representative western blot images of ferroptosis-related proteins (SLC7A11, GPX4, ACSL4, b-actin). (o) Quantification of relative SLC7A11 expression. (p) Quantification of relative GPX4 expression. (q) Quantification of relative ACSL4 expression. ✶ P < 0.05, ✶ ✶ P < 0.01, ✶ ✶ ✶ P < 0.001, ns: Not significant. Images in (a, c, e, j, k, n) are representative of three independent experiments. Quantitative data in (b, d, f-h, i, l, m, o-q) are presented as mean±SD from three independent experiments. PAQR3: Progestin and adipoQ receptor 3, HCC: Hepatocellular carcinoma, EMT: Epithelial-mesenchymal transition, TBARS: Thiobarbituric acid reactive substance, Oe-NC: Overexpression negative control.

Article Snippet: Lipid peroxidation in HCC cells following the indicated treatments was assessed using a TBARS Assay Kit (Elabscience Biotechnology Co., Ltd., E-BC-K298-M, Wuhan, Hubei, China).

Techniques: Wound Healing Assay, Migration, Transwell Invasion Assay, Transwell Assay, Western Blot, Expressing, TBARS Assay, Staining, Fluorescence, Over Expression, Negative Control