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pcna pc10 mouse antibody  (Thermo Fisher)


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    Structured Review

    Thermo Fisher pcna pc10 mouse antibody
    Pcna Pc10 Mouse Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/pcna+pc10+mouse+antibody/pcna+antibody/pmc12108426-34-8-2
    Average 90 stars, based on 1 article reviews
    pcna pc10 mouse antibody - by Bioz Stars, 2026-09
    90/100 stars

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    other:

    Article Title: Immunohistochemical Detection of Iron-Related Proteins in Sertoli Cell-Only Patterns in Canine Testicular Lesions
    Article Snippet: PCNA , ThermoFisher, Carlsbad, CA, USA PC10 , Mouse , 1:400.



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    Fig. 3 H&E (A–B) and Trichrome AZAN (C–D) stainings of representative endometriotic lesions in DCI-treated mice 28 days p.t. by LM. A, D mag. 10X; bar: 100 µm. B, E mag. 20X; bar: 50 µm. C, F mag. 40X; bar: 20 µm (E–F) and Trichrome AZAN (G, H) stainings of representative endometriotic lesions in DG + DCI-treated mice 28 days p.t. by LM. E, G mag. 20X; bar: 50 µm. F, H mag. 40X; bar: 20 µm. H&E (I–K) and Trichrome AZAN (J–L) stainings of representative endometriotic lesions in DG-treated mice 28 days p.t. by LM. I, J mag. 20X; bar: 50 µm. K, L mag. 40X; bar: 20 µm. M–T <t>PCNA</t> immunoreactivity in endometriotic lesions 28 days p.t. of control EMS (M, N), DCI-treated (O, P), DG + DCI (Q, R) and DG-treated (S, T)-treated mice by LM. M, O, Q, S LM, mag. 20X; bar: 50 µm. N, P, R, T mag. 40X; bar: 20 µm. U–AB CD34 immunoreactivity in endometriotic lesions 28 days p.t. of control EMS (U, V), DCI-treated (W, X), DG + DCI (Y, Z) and DG-treated (AA, AB)-treated mice by LM. U, W, Y, AA mag. 20X; bar: 50 µm. V, X, Z, AB mag. 40X; bar: 20 µm
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    Santa Cruz Biotechnology mouse primary antibodies against pcna pc10
    Figure 1. Nuclear distribution of <t>PCNA</t> and RPA1 during hydroxyurea-induced replication stress. (A) Comparison of the expression level of BAC-tagged PCNA-mCherry and RPA1-EGFP versus their endogenous counterparts via Western blotting. (B) Representative timelapse airyscan images of RPA1-EGFP and PCNA-mCherry before and during HU-induced replication stress. Scale bar = 5 µm. (C) Same as (B), but in the presence of ATR inhibitor AZD6738 before and during HU treatment. Scale bar = 5 µm. Abbreviations: HU: hydroxyurea; AZD: AZD6738.
    Mouse Primary Antibodies Against Pcna Pc10, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Fig. 3 H&E (A–B) and Trichrome AZAN (C–D) stainings of representative endometriotic lesions in DCI-treated mice 28 days p.t. by LM. A, D mag. 10X; bar: 100 µm. B, E mag. 20X; bar: 50 µm. C, F mag. 40X; bar: 20 µm (E–F) and Trichrome AZAN (G, H) stainings of representative endometriotic lesions in DG + DCI-treated mice 28 days p.t. by LM. E, G mag. 20X; bar: 50 µm. F, H mag. 40X; bar: 20 µm. H&E (I–K) and Trichrome AZAN (J–L) stainings of representative endometriotic lesions in DG-treated mice 28 days p.t. by LM. I, J mag. 20X; bar: 50 µm. K, L mag. 40X; bar: 20 µm. M–T PCNA immunoreactivity in endometriotic lesions 28 days p.t. of control EMS (M, N), DCI-treated (O, P), DG + DCI (Q, R) and DG-treated (S, T)-treated mice by LM. M, O, Q, S LM, mag. 20X; bar: 50 µm. N, P, R, T mag. 40X; bar: 20 µm. U–AB CD34 immunoreactivity in endometriotic lesions 28 days p.t. of control EMS (U, V), DCI-treated (W, X), DG + DCI (Y, Z) and DG-treated (AA, AB)-treated mice by LM. U, W, Y, AA mag. 20X; bar: 50 µm. V, X, Z, AB mag. 40X; bar: 20 µm

    Journal: Molecular medicine (Cambridge, Mass.)

    Article Title: D-chiro-inositol effectively counteracts endometriosis in a mouse model.

    doi: 10.1186/s10020-025-01178-6

    Figure Lengend Snippet: Fig. 3 H&E (A–B) and Trichrome AZAN (C–D) stainings of representative endometriotic lesions in DCI-treated mice 28 days p.t. by LM. A, D mag. 10X; bar: 100 µm. B, E mag. 20X; bar: 50 µm. C, F mag. 40X; bar: 20 µm (E–F) and Trichrome AZAN (G, H) stainings of representative endometriotic lesions in DG + DCI-treated mice 28 days p.t. by LM. E, G mag. 20X; bar: 50 µm. F, H mag. 40X; bar: 20 µm. H&E (I–K) and Trichrome AZAN (J–L) stainings of representative endometriotic lesions in DG-treated mice 28 days p.t. by LM. I, J mag. 20X; bar: 50 µm. K, L mag. 40X; bar: 20 µm. M–T PCNA immunoreactivity in endometriotic lesions 28 days p.t. of control EMS (M, N), DCI-treated (O, P), DG + DCI (Q, R) and DG-treated (S, T)-treated mice by LM. M, O, Q, S LM, mag. 20X; bar: 50 µm. N, P, R, T mag. 40X; bar: 20 µm. U–AB CD34 immunoreactivity in endometriotic lesions 28 days p.t. of control EMS (U, V), DCI-treated (W, X), DG + DCI (Y, Z) and DG-treated (AA, AB)-treated mice by LM. U, W, Y, AA mag. 20X; bar: 50 µm. V, X, Z, AB mag. 40X; bar: 20 µm

    Article Snippet: For PCNA staining, after thorough washing, sections were incubated with M.O.M mouse IgG blocking reagent (Vector Laboratories Burlingame, CA, USA) overnight at 4 °C according to the manufacturer’s protocol and then incubated with mouse monoclonal anti-PCNA antibody (1:500, NB500-106, Novus Biological) (Mishra et al. 2020) diluted in mouse on mouse (M.O.M) diluent for 30 min, according to the Vector Laboratories instructions.

    Techniques: Control

    Fig. 6 A–D CD34 immunoreactivity in endometriotic ovary 28 days p.t. of control EMS (A), DCI (B), DG + DCI (C) and DG (D)-treated mice by LM. mag. 20X; bar: 50 µm; E–H) PCNA immunoreactivity in endometriotic ovary 28 days p.t. of control EMS (E), DCI (F), DG + DCI (G) and DG (H)-treated mice by LM. mag. 20X; bar: 50 µm. I–L IL-1β immunoreactivity in endometriotic ovary 28 days p.t. of control EMS (I), DCI (J), DG + DCI (K) and DG-treated (L)-treated mice by LM. mag. 20X; bar: 50 µm. M–P E-Cadherin immunoreactivity in endometriotic ovary 28 days p.t. of control EMS (M), DCI (N), DG + DCI (O) and DG (P)-treated mice by LM. mag. 20X; bar: 50 µm

    Journal: Molecular medicine (Cambridge, Mass.)

    Article Title: D-chiro-inositol effectively counteracts endometriosis in a mouse model.

    doi: 10.1186/s10020-025-01178-6

    Figure Lengend Snippet: Fig. 6 A–D CD34 immunoreactivity in endometriotic ovary 28 days p.t. of control EMS (A), DCI (B), DG + DCI (C) and DG (D)-treated mice by LM. mag. 20X; bar: 50 µm; E–H) PCNA immunoreactivity in endometriotic ovary 28 days p.t. of control EMS (E), DCI (F), DG + DCI (G) and DG (H)-treated mice by LM. mag. 20X; bar: 50 µm. I–L IL-1β immunoreactivity in endometriotic ovary 28 days p.t. of control EMS (I), DCI (J), DG + DCI (K) and DG-treated (L)-treated mice by LM. mag. 20X; bar: 50 µm. M–P E-Cadherin immunoreactivity in endometriotic ovary 28 days p.t. of control EMS (M), DCI (N), DG + DCI (O) and DG (P)-treated mice by LM. mag. 20X; bar: 50 µm

    Article Snippet: For PCNA staining, after thorough washing, sections were incubated with M.O.M mouse IgG blocking reagent (Vector Laboratories Burlingame, CA, USA) overnight at 4 °C according to the manufacturer’s protocol and then incubated with mouse monoclonal anti-PCNA antibody (1:500, NB500-106, Novus Biological) (Mishra et al. 2020) diluted in mouse on mouse (M.O.M) diluent for 30 min, according to the Vector Laboratories instructions.

    Techniques: Control

    Figure 1. Nuclear distribution of PCNA and RPA1 during hydroxyurea-induced replication stress. (A) Comparison of the expression level of BAC-tagged PCNA-mCherry and RPA1-EGFP versus their endogenous counterparts via Western blotting. (B) Representative timelapse airyscan images of RPA1-EGFP and PCNA-mCherry before and during HU-induced replication stress. Scale bar = 5 µm. (C) Same as (B), but in the presence of ATR inhibitor AZD6738 before and during HU treatment. Scale bar = 5 µm. Abbreviations: HU: hydroxyurea; AZD: AZD6738.

    Journal: International journal of molecular sciences

    Article Title: In and out of Replication Stress: PCNA/RPA1-Based Dynamics of Fork Stalling and Restart in the Same Cell.

    doi: 10.3390/ijms26020667

    Figure Lengend Snippet: Figure 1. Nuclear distribution of PCNA and RPA1 during hydroxyurea-induced replication stress. (A) Comparison of the expression level of BAC-tagged PCNA-mCherry and RPA1-EGFP versus their endogenous counterparts via Western blotting. (B) Representative timelapse airyscan images of RPA1-EGFP and PCNA-mCherry before and during HU-induced replication stress. Scale bar = 5 µm. (C) Same as (B), but in the presence of ATR inhibitor AZD6738 before and during HU treatment. Scale bar = 5 µm. Abbreviations: HU: hydroxyurea; AZD: AZD6738.

    Article Snippet: After blocking, membranes were incubated with mouse primary antibodies against PCNA (PC10) (sc-56, Santa Cruz Biotechnology, Dallas, TX, USA) and RPA70/RPA1 (MBS600132, MyBioSource, San Diego, CA, USA) overnight at 4 ◦C, followed by incubation with an IRDye® 800CW Goat anti-Mouse IgG Secondary Antibody (926-32210, LICORbio, Lincoln, NE, USA) at room temperature for 1 h. Bands were visualized using an Odyssey Infrared Imaging System (LICORbio) and saved as TIF images.

    Techniques: Comparison, Expressing, Western Blot

    Figure 2. Dynamics of RPA1 and PCNA during HU-induced replication fork stalling and restart. (A) Representative time-lapse images of RPA1 and PCNA before, during, and after HU treatment. Arrows indicate timepoints of HU addition and washout. Scale bar = 5 µm. (B) Same as (A), but with inhibition of ATR (3 µM AZD6738) throughout the experimental period. (C) Normalized intensity of PCNA and RPA1 at replication foci during HU-induced replication fork stalling and restart, with or without ATR inhibition. The maximum intensity of PCNA/RPA at replication foci is normalized to 1. (D) Fraction of PCNA and RPA1 bound at replication foci during HU-induced replication fork stalling and restart, with or without ATR inhibition, relative to the total nuclear intensity of PCNA/RPA1, which is normalized to 1. (E) Estimated number of PCNA homotrimer and RPA heterotrimer complexes engaged at replication foci with or without ATR inhibition. (F) Estimated number of nucleotides covered by RPA heterotrimers with or without ATR inhibition. For HU only: n = 17 cells; for HU + AZD: n = 10 cells. Abbreviations: HU: hydroxyurea; AZD: AZD6738.

    Journal: International journal of molecular sciences

    Article Title: In and out of Replication Stress: PCNA/RPA1-Based Dynamics of Fork Stalling and Restart in the Same Cell.

    doi: 10.3390/ijms26020667

    Figure Lengend Snippet: Figure 2. Dynamics of RPA1 and PCNA during HU-induced replication fork stalling and restart. (A) Representative time-lapse images of RPA1 and PCNA before, during, and after HU treatment. Arrows indicate timepoints of HU addition and washout. Scale bar = 5 µm. (B) Same as (A), but with inhibition of ATR (3 µM AZD6738) throughout the experimental period. (C) Normalized intensity of PCNA and RPA1 at replication foci during HU-induced replication fork stalling and restart, with or without ATR inhibition. The maximum intensity of PCNA/RPA at replication foci is normalized to 1. (D) Fraction of PCNA and RPA1 bound at replication foci during HU-induced replication fork stalling and restart, with or without ATR inhibition, relative to the total nuclear intensity of PCNA/RPA1, which is normalized to 1. (E) Estimated number of PCNA homotrimer and RPA heterotrimer complexes engaged at replication foci with or without ATR inhibition. (F) Estimated number of nucleotides covered by RPA heterotrimers with or without ATR inhibition. For HU only: n = 17 cells; for HU + AZD: n = 10 cells. Abbreviations: HU: hydroxyurea; AZD: AZD6738.

    Article Snippet: After blocking, membranes were incubated with mouse primary antibodies against PCNA (PC10) (sc-56, Santa Cruz Biotechnology, Dallas, TX, USA) and RPA70/RPA1 (MBS600132, MyBioSource, San Diego, CA, USA) overnight at 4 ◦C, followed by incubation with an IRDye® 800CW Goat anti-Mouse IgG Secondary Antibody (926-32210, LICORbio, Lincoln, NE, USA) at room temperature for 1 h. Bands were visualized using an Odyssey Infrared Imaging System (LICORbio) and saved as TIF images.

    Techniques: Inhibition

    Figure 3. Detailed overview for the measurement and quantification of PCNA and RPA1 engaged at replication foci during fork stalling and restart. (A) Regions of interest applied for analysis. ‘A’ represents the cell nucleus (white lining), ‘B’ represents a region within the nucleus without replication foci (blue ellipse), and ‘C’ represents noise outside of cells (orange square). (B) Constants and formulas for the calculation of variables used for quantifying RPA1 and PCNA kinetics. (C) Mean intensity of diffusing RPA1-EGFP/mPCNA-mCherry within region ‘B’, calculated as shown in the formula for Dt.

    Journal: International journal of molecular sciences

    Article Title: In and out of Replication Stress: PCNA/RPA1-Based Dynamics of Fork Stalling and Restart in the Same Cell.

    doi: 10.3390/ijms26020667

    Figure Lengend Snippet: Figure 3. Detailed overview for the measurement and quantification of PCNA and RPA1 engaged at replication foci during fork stalling and restart. (A) Regions of interest applied for analysis. ‘A’ represents the cell nucleus (white lining), ‘B’ represents a region within the nucleus without replication foci (blue ellipse), and ‘C’ represents noise outside of cells (orange square). (B) Constants and formulas for the calculation of variables used for quantifying RPA1 and PCNA kinetics. (C) Mean intensity of diffusing RPA1-EGFP/mPCNA-mCherry within region ‘B’, calculated as shown in the formula for Dt.

    Article Snippet: After blocking, membranes were incubated with mouse primary antibodies against PCNA (PC10) (sc-56, Santa Cruz Biotechnology, Dallas, TX, USA) and RPA70/RPA1 (MBS600132, MyBioSource, San Diego, CA, USA) overnight at 4 ◦C, followed by incubation with an IRDye® 800CW Goat anti-Mouse IgG Secondary Antibody (926-32210, LICORbio, Lincoln, NE, USA) at room temperature for 1 h. Bands were visualized using an Odyssey Infrared Imaging System (LICORbio) and saved as TIF images.

    Techniques:

    Figure 4. Exchange of PCNA and RPA1 at replication foci during unperturbed and stalled replication. (A) Representative timelapse images depicting simultaneous FRAP of RPA1 and residual PCNA co-localized at stalled replication forks under nucleotide depletion. (B) Same as (A), but under conditions of ATR inhibition. (C) Replication timelapse images of PCNA FRAP at replication foci during unperturbed replication (upper panel) or in the presence of ATR inhibitor AZD6738. (D) FRAP curves of PCNA under the following conditions: untreated, HU alone, AZD alone, HU + AZD. The mean intensity is normalized as described in Figure S5. (E) Contribution of distinct PCNA fractions (freely diffusing [F1] and replisome-bound [F2]) to the FRAP curve under nucleotide depletion, as determined via fitting of two single exponential curves. (F) Same as (E), but in the presence of both AZD6738 and HU. (G) Recovery of the replisome-bound fraction of PCNA under HU treatment, as derived based on (E). (H) Recovery of the replisome-bound fraction of PCNA under HU treatment, as derived based on (F). (I) Comparison of PCNA recovery under the following conditions: untreated, AZD alone, HU alone, HU + AZD. The contribution of freely diffusing PCNA has been subtracted from the HU and HU + AZD curves, as per (G,H). (J) Number of PCNA complexes at a single replication fork, recovered after photobleaching. (K) Enlarged view of HU and HU + AZD curves from (J). (L) FRAP curves of RPA1 at replication foci under HU alone and HU + AZD. (M) Number of RPA complexes at a single replication fork recovered after photobleaching under HU alone and HU + AZD. For HU: n = 11 cells; for HU + AZD: n = 16 cells; for AZD (PCNA only): n = 13 cells; untreated (PCNA only): n = 15 cells. Abbreviations: HU: hydroxyurea; AZD: AZD6738.

    Journal: International journal of molecular sciences

    Article Title: In and out of Replication Stress: PCNA/RPA1-Based Dynamics of Fork Stalling and Restart in the Same Cell.

    doi: 10.3390/ijms26020667

    Figure Lengend Snippet: Figure 4. Exchange of PCNA and RPA1 at replication foci during unperturbed and stalled replication. (A) Representative timelapse images depicting simultaneous FRAP of RPA1 and residual PCNA co-localized at stalled replication forks under nucleotide depletion. (B) Same as (A), but under conditions of ATR inhibition. (C) Replication timelapse images of PCNA FRAP at replication foci during unperturbed replication (upper panel) or in the presence of ATR inhibitor AZD6738. (D) FRAP curves of PCNA under the following conditions: untreated, HU alone, AZD alone, HU + AZD. The mean intensity is normalized as described in Figure S5. (E) Contribution of distinct PCNA fractions (freely diffusing [F1] and replisome-bound [F2]) to the FRAP curve under nucleotide depletion, as determined via fitting of two single exponential curves. (F) Same as (E), but in the presence of both AZD6738 and HU. (G) Recovery of the replisome-bound fraction of PCNA under HU treatment, as derived based on (E). (H) Recovery of the replisome-bound fraction of PCNA under HU treatment, as derived based on (F). (I) Comparison of PCNA recovery under the following conditions: untreated, AZD alone, HU alone, HU + AZD. The contribution of freely diffusing PCNA has been subtracted from the HU and HU + AZD curves, as per (G,H). (J) Number of PCNA complexes at a single replication fork, recovered after photobleaching. (K) Enlarged view of HU and HU + AZD curves from (J). (L) FRAP curves of RPA1 at replication foci under HU alone and HU + AZD. (M) Number of RPA complexes at a single replication fork recovered after photobleaching under HU alone and HU + AZD. For HU: n = 11 cells; for HU + AZD: n = 16 cells; for AZD (PCNA only): n = 13 cells; untreated (PCNA only): n = 15 cells. Abbreviations: HU: hydroxyurea; AZD: AZD6738.

    Article Snippet: After blocking, membranes were incubated with mouse primary antibodies against PCNA (PC10) (sc-56, Santa Cruz Biotechnology, Dallas, TX, USA) and RPA70/RPA1 (MBS600132, MyBioSource, San Diego, CA, USA) overnight at 4 ◦C, followed by incubation with an IRDye® 800CW Goat anti-Mouse IgG Secondary Antibody (926-32210, LICORbio, Lincoln, NE, USA) at room temperature for 1 h. Bands were visualized using an Odyssey Infrared Imaging System (LICORbio) and saved as TIF images.

    Techniques: Inhibition, Derivative Assay, Comparison

    Figure 5. Influence of ATM activity on RPA1 and PCNA dynamics during HU-induced replication fork stalling and restart under conditions of ATR inhibition. (A) Representative time-lapse images of RPA1 and PCNA before, during, and after 10 mM HU treatment under combined ATM (10 µM Ku55933) and ATR (3 µM AZD6738) inhibition. Arrows indicate timepoints of HU addition and washout. Scale bar = 5 µm. (B) Normalized intensity of PCNA and RPA1 at replication foci during HU-induced replication fork stalling and restart under ATM inhibition with or without ATR co- inhibition. The maximum intensity of PCNA/RPA1 engaged at replication foci is normalized to 1. (C) Fraction of PCNA and RPA1 bound at replication foci during HU-induced replication fork stalling and restart under ATM inhibition with or without ATR inhibition, relative to the total nuclear intensity of PCNA/RPA1, which is normalized to 1. (D) Normalized intensity of PCNA and RPA1 at replication foci during HU-induced replication fork stalling and restart under ATR inhibition with or without ATM co-inhibition. The maximum intensity of PCNA/RPA1 engaged at replication foci is normalized to 1. (E) Fraction of PCNA and RPA1 bound at replication factories during HU-induced replication fork stalling and restart under ATR inhibition with or without ATM inhibition, relative to the total nuclear intensity of PCNA/RPA1, which is normalized to 1. (F) Same as (B,D), but with or without combined ATR + ATM inhibition. (G) Same as (C,E), but with or without combined ATR + ATM inhibition. (H) Estimated number of PCNA homotrimer and RPA heterotrimer complexes engaged at replication foci during HU-induced fork stalling and subsequent restart. (I) Estimated number of nucleotides covered by RPA heterotrimers under combined ATM and ATR inhibition. Dashed orange lines indicate the timepoints of HU addition and wash-out. For HU + AZD + KU, n = 10 cells; for HU + KU: n = 19 cells. Abbreviations: HU: hydroxyurea; AZD: AZD6738; KU: Ku55933.

    Journal: International journal of molecular sciences

    Article Title: In and out of Replication Stress: PCNA/RPA1-Based Dynamics of Fork Stalling and Restart in the Same Cell.

    doi: 10.3390/ijms26020667

    Figure Lengend Snippet: Figure 5. Influence of ATM activity on RPA1 and PCNA dynamics during HU-induced replication fork stalling and restart under conditions of ATR inhibition. (A) Representative time-lapse images of RPA1 and PCNA before, during, and after 10 mM HU treatment under combined ATM (10 µM Ku55933) and ATR (3 µM AZD6738) inhibition. Arrows indicate timepoints of HU addition and washout. Scale bar = 5 µm. (B) Normalized intensity of PCNA and RPA1 at replication foci during HU-induced replication fork stalling and restart under ATM inhibition with or without ATR co- inhibition. The maximum intensity of PCNA/RPA1 engaged at replication foci is normalized to 1. (C) Fraction of PCNA and RPA1 bound at replication foci during HU-induced replication fork stalling and restart under ATM inhibition with or without ATR inhibition, relative to the total nuclear intensity of PCNA/RPA1, which is normalized to 1. (D) Normalized intensity of PCNA and RPA1 at replication foci during HU-induced replication fork stalling and restart under ATR inhibition with or without ATM co-inhibition. The maximum intensity of PCNA/RPA1 engaged at replication foci is normalized to 1. (E) Fraction of PCNA and RPA1 bound at replication factories during HU-induced replication fork stalling and restart under ATR inhibition with or without ATM inhibition, relative to the total nuclear intensity of PCNA/RPA1, which is normalized to 1. (F) Same as (B,D), but with or without combined ATR + ATM inhibition. (G) Same as (C,E), but with or without combined ATR + ATM inhibition. (H) Estimated number of PCNA homotrimer and RPA heterotrimer complexes engaged at replication foci during HU-induced fork stalling and subsequent restart. (I) Estimated number of nucleotides covered by RPA heterotrimers under combined ATM and ATR inhibition. Dashed orange lines indicate the timepoints of HU addition and wash-out. For HU + AZD + KU, n = 10 cells; for HU + KU: n = 19 cells. Abbreviations: HU: hydroxyurea; AZD: AZD6738; KU: Ku55933.

    Article Snippet: After blocking, membranes were incubated with mouse primary antibodies against PCNA (PC10) (sc-56, Santa Cruz Biotechnology, Dallas, TX, USA) and RPA70/RPA1 (MBS600132, MyBioSource, San Diego, CA, USA) overnight at 4 ◦C, followed by incubation with an IRDye® 800CW Goat anti-Mouse IgG Secondary Antibody (926-32210, LICORbio, Lincoln, NE, USA) at room temperature for 1 h. Bands were visualized using an Odyssey Infrared Imaging System (LICORbio) and saved as TIF images.

    Techniques: Activity Assay, Inhibition

    Figure 6. Dynamics of PCNA and RPA1 during hydroxyurea-induced replication stress in HeLa, DU145, and PC3 cell lines. (A) Normalized intensity of PCNA and RPA1 during, before, and after HU treatment. (B) Same as (A), but under conditions of ATM inhibition (10 µM Ku55933). (C) Same as (A), but under conditions of ATR inhibition (3 µM AZD6738). (D) Same as (A), but under combined ATR and ATM inhibition. Dashed green lines indicate the timepoints of HU addition and washout. Data are presented as the mean ± SD. For HeLa: n = 11 cells (HU), n = 16 cells (HU + AZD), n = 19 cells (HU + KU), n = 10 cells (HU + AZD + KU); for DU145: n = 11 cells (HU), n = 10 cells (HU + AZD), n = 10 cells (HU + KU), n = 18 cells (HU + AZD + KU); for PC3: n = 18 cells (HU), n = 10 cells (HU + AZD), n = 13 cells (HU + KU), n = 15 cells (HU + AZD + KU). Abbreviations: HU: hydroxyurea; AZD: AZD6738; KU: Ku55933.

    Journal: International journal of molecular sciences

    Article Title: In and out of Replication Stress: PCNA/RPA1-Based Dynamics of Fork Stalling and Restart in the Same Cell.

    doi: 10.3390/ijms26020667

    Figure Lengend Snippet: Figure 6. Dynamics of PCNA and RPA1 during hydroxyurea-induced replication stress in HeLa, DU145, and PC3 cell lines. (A) Normalized intensity of PCNA and RPA1 during, before, and after HU treatment. (B) Same as (A), but under conditions of ATM inhibition (10 µM Ku55933). (C) Same as (A), but under conditions of ATR inhibition (3 µM AZD6738). (D) Same as (A), but under combined ATR and ATM inhibition. Dashed green lines indicate the timepoints of HU addition and washout. Data are presented as the mean ± SD. For HeLa: n = 11 cells (HU), n = 16 cells (HU + AZD), n = 19 cells (HU + KU), n = 10 cells (HU + AZD + KU); for DU145: n = 11 cells (HU), n = 10 cells (HU + AZD), n = 10 cells (HU + KU), n = 18 cells (HU + AZD + KU); for PC3: n = 18 cells (HU), n = 10 cells (HU + AZD), n = 13 cells (HU + KU), n = 15 cells (HU + AZD + KU). Abbreviations: HU: hydroxyurea; AZD: AZD6738; KU: Ku55933.

    Article Snippet: After blocking, membranes were incubated with mouse primary antibodies against PCNA (PC10) (sc-56, Santa Cruz Biotechnology, Dallas, TX, USA) and RPA70/RPA1 (MBS600132, MyBioSource, San Diego, CA, USA) overnight at 4 ◦C, followed by incubation with an IRDye® 800CW Goat anti-Mouse IgG Secondary Antibody (926-32210, LICORbio, Lincoln, NE, USA) at room temperature for 1 h. Bands were visualized using an Odyssey Infrared Imaging System (LICORbio) and saved as TIF images.

    Techniques: Inhibition

    Figure 7. Dynamics of RAD18 during hydroxyurea-induced replication fork stalling and restart. (A) Representative time-lapse images of RAD18 and PCNA before, during, and after HU treatment. Arrows indicate timepoints of HU addition and washout. Scale bar = 5 µm. (B) Normalized intensity of RAD18 and PCNA during HU-induced replication fork stalling and restart, with or without ATR inhibition (3 µM AZD6738). The maximum intensity of PCNA/RAD18 foci is normalized to 1. For HU: n = 17 cells; for HU + AZD: n = 12 cells. (C) Normalized intensity of single RAD18 and PCNA foci before and after HU addition (max intensity = 1). Single foci were tracked using SPARTACUSS, and a representative kymogram is shown. This panel presents a case where RAD18 accumulates at a replication focus, while PCNA dissociates after HU treatment. (D) Same as (C), but under conditions of ATR inhibition. (E) Same as (C), but this panel presents a case where RAD18 is already present at the replication focus and dissociates from the replication focus in parallel to PCNA upon HU addition. (F) Same scenario as shown in (E), but under conditions of ATR inhibition. Dashed green lines indicate timepoints of HU addition and removal. Abbreviations: HU: hydroxyurea; AZD: AZD6738.

    Journal: International journal of molecular sciences

    Article Title: In and out of Replication Stress: PCNA/RPA1-Based Dynamics of Fork Stalling and Restart in the Same Cell.

    doi: 10.3390/ijms26020667

    Figure Lengend Snippet: Figure 7. Dynamics of RAD18 during hydroxyurea-induced replication fork stalling and restart. (A) Representative time-lapse images of RAD18 and PCNA before, during, and after HU treatment. Arrows indicate timepoints of HU addition and washout. Scale bar = 5 µm. (B) Normalized intensity of RAD18 and PCNA during HU-induced replication fork stalling and restart, with or without ATR inhibition (3 µM AZD6738). The maximum intensity of PCNA/RAD18 foci is normalized to 1. For HU: n = 17 cells; for HU + AZD: n = 12 cells. (C) Normalized intensity of single RAD18 and PCNA foci before and after HU addition (max intensity = 1). Single foci were tracked using SPARTACUSS, and a representative kymogram is shown. This panel presents a case where RAD18 accumulates at a replication focus, while PCNA dissociates after HU treatment. (D) Same as (C), but under conditions of ATR inhibition. (E) Same as (C), but this panel presents a case where RAD18 is already present at the replication focus and dissociates from the replication focus in parallel to PCNA upon HU addition. (F) Same scenario as shown in (E), but under conditions of ATR inhibition. Dashed green lines indicate timepoints of HU addition and removal. Abbreviations: HU: hydroxyurea; AZD: AZD6738.

    Article Snippet: After blocking, membranes were incubated with mouse primary antibodies against PCNA (PC10) (sc-56, Santa Cruz Biotechnology, Dallas, TX, USA) and RPA70/RPA1 (MBS600132, MyBioSource, San Diego, CA, USA) overnight at 4 ◦C, followed by incubation with an IRDye® 800CW Goat anti-Mouse IgG Secondary Antibody (926-32210, LICORbio, Lincoln, NE, USA) at room temperature for 1 h. Bands were visualized using an Odyssey Infrared Imaging System (LICORbio) and saved as TIF images.

    Techniques: Inhibition

    Figure 8. Visual summary of PCNA and RPA dynamics during replication fork stalling and restart.

    Journal: International journal of molecular sciences

    Article Title: In and out of Replication Stress: PCNA/RPA1-Based Dynamics of Fork Stalling and Restart in the Same Cell.

    doi: 10.3390/ijms26020667

    Figure Lengend Snippet: Figure 8. Visual summary of PCNA and RPA dynamics during replication fork stalling and restart.

    Article Snippet: After blocking, membranes were incubated with mouse primary antibodies against PCNA (PC10) (sc-56, Santa Cruz Biotechnology, Dallas, TX, USA) and RPA70/RPA1 (MBS600132, MyBioSource, San Diego, CA, USA) overnight at 4 ◦C, followed by incubation with an IRDye® 800CW Goat anti-Mouse IgG Secondary Antibody (926-32210, LICORbio, Lincoln, NE, USA) at room temperature for 1 h. Bands were visualized using an Odyssey Infrared Imaging System (LICORbio) and saved as TIF images.

    Techniques: