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Exposure of preeclamptic pMSCs to Hinokitiol rescues FNglycosylation and matrix deposition. (A) Western blot for FN and Con A following immunoprecipitation of FN in TC and PE pMSCs. n = 3 separate pMSC isolations. (B) Intracellular Fe 2+ content in TC and PE pMSCs. n = 4 separate pMSC isolations performed in duplicate. (C) Western blot and densitometry for FN in PE pMSCs upon exposure to 1 μM Hinokitiol. n = 4 separate pMSC isolations. (D) Western blot for FN and Con A following immunoprecipitation of FN in PE pMSCs exposed to 1 μM Hinokitiol. Representative of two separate pMSC isolations. (E) IF for FN (green) and the organelle markers <t>CALR</t> (ER), STX6 (Golgi), and LAMP1 (lysosomes) in red (top, middle, and bottom panels, respectively) following 1 μM Hinokitiol exposure. In IF images, arrowheads indicate FN fibrillar deposition while stars show FN clustering. Nuclei were counterstained with DAPI. Scale bars at 63× magnification represent 7 μM. Data are presented as mean ± SEM. * P ≤ 0.05 (Unpaired Student’s t -test).
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Exposure of preeclamptic pMSCs to Hinokitiol rescues FNglycosylation and matrix deposition. (A) Western blot for FN and Con A following immunoprecipitation of FN in TC and PE pMSCs. n = 3 separate pMSC isolations. (B) Intracellular Fe 2+ content in TC and PE pMSCs. n = 4 separate pMSC isolations performed in duplicate. (C) Western blot and densitometry for FN in PE pMSCs upon exposure to 1 μM Hinokitiol. n = 4 separate pMSC isolations. (D) Western blot for FN and Con A following immunoprecipitation of FN in PE pMSCs exposed to 1 μM Hinokitiol. Representative of two separate pMSC isolations. (E) IF for FN (green) and the organelle markers <t>CALR</t> (ER), STX6 (Golgi), and LAMP1 (lysosomes) in red (top, middle, and bottom panels, respectively) following 1 μM Hinokitiol exposure. In IF images, arrowheads indicate FN fibrillar deposition while stars show FN clustering. Nuclei were counterstained with DAPI. Scale bars at 63× magnification represent 7 μM. Data are presented as mean ± SEM. * P ≤ 0.05 (Unpaired Student’s t -test).
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Image Search Results


Exposure of preeclamptic pMSCs to Hinokitiol rescues FNglycosylation and matrix deposition. (A) Western blot for FN and Con A following immunoprecipitation of FN in TC and PE pMSCs. n = 3 separate pMSC isolations. (B) Intracellular Fe 2+ content in TC and PE pMSCs. n = 4 separate pMSC isolations performed in duplicate. (C) Western blot and densitometry for FN in PE pMSCs upon exposure to 1 μM Hinokitiol. n = 4 separate pMSC isolations. (D) Western blot for FN and Con A following immunoprecipitation of FN in PE pMSCs exposed to 1 μM Hinokitiol. Representative of two separate pMSC isolations. (E) IF for FN (green) and the organelle markers CALR (ER), STX6 (Golgi), and LAMP1 (lysosomes) in red (top, middle, and bottom panels, respectively) following 1 μM Hinokitiol exposure. In IF images, arrowheads indicate FN fibrillar deposition while stars show FN clustering. Nuclei were counterstained with DAPI. Scale bars at 63× magnification represent 7 μM. Data are presented as mean ± SEM. * P ≤ 0.05 (Unpaired Student’s t -test).

Journal: Frontiers in Cell and Developmental Biology

Article Title: JMJD6 Dysfunction Due to Iron Deficiency in Preeclampsia Disrupts Fibronectin Homeostasis Resulting in Diminished Trophoblast Migration

doi: 10.3389/fcell.2021.652607

Figure Lengend Snippet: Exposure of preeclamptic pMSCs to Hinokitiol rescues FNglycosylation and matrix deposition. (A) Western blot for FN and Con A following immunoprecipitation of FN in TC and PE pMSCs. n = 3 separate pMSC isolations. (B) Intracellular Fe 2+ content in TC and PE pMSCs. n = 4 separate pMSC isolations performed in duplicate. (C) Western blot and densitometry for FN in PE pMSCs upon exposure to 1 μM Hinokitiol. n = 4 separate pMSC isolations. (D) Western blot for FN and Con A following immunoprecipitation of FN in PE pMSCs exposed to 1 μM Hinokitiol. Representative of two separate pMSC isolations. (E) IF for FN (green) and the organelle markers CALR (ER), STX6 (Golgi), and LAMP1 (lysosomes) in red (top, middle, and bottom panels, respectively) following 1 μM Hinokitiol exposure. In IF images, arrowheads indicate FN fibrillar deposition while stars show FN clustering. Nuclei were counterstained with DAPI. Scale bars at 63× magnification represent 7 μM. Data are presented as mean ± SEM. * P ≤ 0.05 (Unpaired Student’s t -test).

Article Snippet: Primary antibodies employed in IF and WB include mouse monoclonal anti-β-actin (ACTB) (sc-477778, 1:1,000, Santa Cruz), mouse monoclonal anti-integrin α5β1 (MAB1999, 1:200, Millipore Sigma), mouse monoclonal anti-α-tubulin (sc-8035, 1:1,000, Santa Cruz), mouse monoclonal anti-Calreticulin (CALR) (ab22683, 1:300, Santa Cruz Biotechnology), mouse monoclonal anti-CK7 (sc-70936, 1:200, Santa Cruz), rabbit polyclonal anti-collagen IV (COL IV) (ab19808, 1:200, Abcam), mouse monoclonal anti-Fibronectin (sc-8422, 1:1,000, Santa Cruz Biotechnology), rabbit polyclonal anti-Fibronectin (ab2413, 1:2,000, Abcam), mouse monoclonal anti-HLAG (ab52454, 1:200, Abcam), mouse monoclonal anti-JMJD6 (sc-28348, 1:500, Santa Cruz), rabbit polyclonal anti-JMJD6 (ab64575, 1:1,000, Abcam), rabbit monoclonal anti-Syntaxin 6 (CB4B2 #2869.

Techniques: Western Blot, Immunoprecipitation

Iron supplementation of PE pMSCs improves FN deposition. (A) Western blot and corresponding densitometry for FN in TC pMSCs (left panels) and PE pMSCs (right panels) treated with 25 or 100 μM FAC. n ≥ 3 separate pMSC isolations per group performed in duplicate. (B) IF for FN (green) and CALR (red) in TC pMSCs exposed to 50 μM of the iron chelator, DFO (left panel), and in PE pMSCs exposed to 100 μM FAC (right panel). Scale bars at 63× magnification represent 7 μM. Representative of three separate pMSC isolations per group. (C) Quantification of the labile iron pool in TC pMSCs exposed to 50 μM DFO or 100 μM FAC (left panel) and in PE pMSCs exposed to 100 μM FAC and 1 μM Hinokitiol (right panel). Values are expressed as fold change of relative fluorescence units (RFU; measured at 488 nM excitation/517 nM emission) over vehicle controls. n = 4 and 3 separate pMSC isolations, respectively, per group. Data are presented as mean ± SEM. * P ≤ 0.05, ** P ≤ 0.01 (one-way ANOVA and Bonferroni post-test ).

Journal: Frontiers in Cell and Developmental Biology

Article Title: JMJD6 Dysfunction Due to Iron Deficiency in Preeclampsia Disrupts Fibronectin Homeostasis Resulting in Diminished Trophoblast Migration

doi: 10.3389/fcell.2021.652607

Figure Lengend Snippet: Iron supplementation of PE pMSCs improves FN deposition. (A) Western blot and corresponding densitometry for FN in TC pMSCs (left panels) and PE pMSCs (right panels) treated with 25 or 100 μM FAC. n ≥ 3 separate pMSC isolations per group performed in duplicate. (B) IF for FN (green) and CALR (red) in TC pMSCs exposed to 50 μM of the iron chelator, DFO (left panel), and in PE pMSCs exposed to 100 μM FAC (right panel). Scale bars at 63× magnification represent 7 μM. Representative of three separate pMSC isolations per group. (C) Quantification of the labile iron pool in TC pMSCs exposed to 50 μM DFO or 100 μM FAC (left panel) and in PE pMSCs exposed to 100 μM FAC and 1 μM Hinokitiol (right panel). Values are expressed as fold change of relative fluorescence units (RFU; measured at 488 nM excitation/517 nM emission) over vehicle controls. n = 4 and 3 separate pMSC isolations, respectively, per group. Data are presented as mean ± SEM. * P ≤ 0.05, ** P ≤ 0.01 (one-way ANOVA and Bonferroni post-test ).

Article Snippet: Primary antibodies employed in IF and WB include mouse monoclonal anti-β-actin (ACTB) (sc-477778, 1:1,000, Santa Cruz), mouse monoclonal anti-integrin α5β1 (MAB1999, 1:200, Millipore Sigma), mouse monoclonal anti-α-tubulin (sc-8035, 1:1,000, Santa Cruz), mouse monoclonal anti-Calreticulin (CALR) (ab22683, 1:300, Santa Cruz Biotechnology), mouse monoclonal anti-CK7 (sc-70936, 1:200, Santa Cruz), rabbit polyclonal anti-collagen IV (COL IV) (ab19808, 1:200, Abcam), mouse monoclonal anti-Fibronectin (sc-8422, 1:1,000, Santa Cruz Biotechnology), rabbit polyclonal anti-Fibronectin (ab2413, 1:2,000, Abcam), mouse monoclonal anti-HLAG (ab52454, 1:200, Abcam), mouse monoclonal anti-JMJD6 (sc-28348, 1:500, Santa Cruz), rabbit polyclonal anti-JMJD6 (ab64575, 1:1,000, Abcam), rabbit monoclonal anti-Syntaxin 6 (CB4B2 #2869.

Techniques: Western Blot, Fluorescence