Journal: bioRxiv

Article Title: Conditional deletion of human STN1 leads to telomere dysfunction and telomerase-dependent genome instability and proliferation defects

doi: 10.1101/2025.08.06.669015

Figure Lengend Snippet: (A) Western blot of STN1 levels in HeLa iCas9 sgSTN1 cells, as indicated. Actinin serves as a loading control. DOX=Doxycycline. (B) Growth curve analysis of STN1 KO and control cells. D=day after DOX addition. n=3 independent, biological replicates. (C-D) Flow cytometry analysis of STN1 KO cells. (C) Representative histograms of STN1 KO and control (-DOX) cells. Left, histograms of DNA content versus cell count. Right, DNA content versus EdU intensity. (D) Percentage of G2/M cells over time in STN1 KO cells. n=3 independent, biological replicates. Average values in the graphs indicate the mean and error bars denote ±s.e.m. P -values were calculated by a two-tailed, unpaired t -test.

Article Snippet: Primary: OBFC1 (STN1) (Novus, NBP2-01006), Actinin (Santa Cruz, sc17829), pH3 S10 (Cell Signaling, 9706), RPA32 (Abcam, ab16850), and γH2AX (Bethyl, A300-081A; Abcam, ab81299).

Techniques: Western Blot, Control, Flow Cytometry, Cell Counting, Two Tailed Test

Journal: bioRxiv

Article Title: Conditional deletion of human STN1 leads to telomere dysfunction and telomerase-dependent genome instability and proliferation defects

doi: 10.1101/2025.08.06.669015

Figure Lengend Snippet: (A) Representative image of RPA-γH2AX foci. STN1 KO image is from day 12 after DOX addition. (B-C) Analysis of foci in STN1 KO and control (-DOX) cells. Percentage of nuclei displaying 3 or more RPA (B) or RPA-γH2AX foci (C). n=3 independent, biological replicates. Average values in the graphs indicate the mean and error bars denote ±s.e.m. P -values were calculated by a two-tailed, unpaired t -test. DOX=Doxycycline. D=day after DOX addition.

Article Snippet: Primary: OBFC1 (STN1) (Novus, NBP2-01006), Actinin (Santa Cruz, sc17829), pH3 S10 (Cell Signaling, 9706), RPA32 (Abcam, ab16850), and γH2AX (Bethyl, A300-081A; Abcam, ab81299).

Techniques: Control, Two Tailed Test

Journal: bioRxiv

Article Title: Conditional deletion of human STN1 leads to telomere dysfunction and telomerase-dependent genome instability and proliferation defects

doi: 10.1101/2025.08.06.669015

Figure Lengend Snippet: (A) Western blot of STN1 and Flag-STN1 levels, as indicated. KO+Flag-STN1 denotes STN1 KO cells expressing Flag-STN1. Actinin serves as a loading control. (B) Percentage of nuclei displaying 3 or more RPA foci on day 15 after DOX addition. n=3 independent, biological replicates. (C) Growth curve analysis, as indicated. n=3 independent, biological replicates. (D) Percentage of G2/M cells determined by flow cytometry on day 15 after DOX addition. n=4 independent, biological replicates. Average values in the graphs indicate the mean and error bars denote ±s.e.m. P -values were calculated by a two-tailed, unpaired t -test. DOX=Doxycycline.

Article Snippet: Primary: OBFC1 (STN1) (Novus, NBP2-01006), Actinin (Santa Cruz, sc17829), pH3 S10 (Cell Signaling, 9706), RPA32 (Abcam, ab16850), and γH2AX (Bethyl, A300-081A; Abcam, ab81299).

Techniques: Western Blot, Expressing, Control, Flow Cytometry, Two Tailed Test

Journal: bioRxiv

Article Title: Conditional deletion of human STN1 leads to telomere dysfunction and telomerase-dependent genome instability and proliferation defects

doi: 10.1101/2025.08.06.669015

Figure Lengend Snippet: (A) Representative image of an anaphase bridge (indicated by the arrow) in STN1 KO cells. (B-C) Percentage of anaphase bridges in STN1 KO and control (-DOX) cells between days 5-15 (B) or with expression of exogenous Flag-STN1 on day 15 (C). n=≥3 independent, biological replicates. (D) Representative image of micronuclei (indicated by the arrows) in the STN1 KO cells. (E-F) Percentage of cells with micronuclei in STN1 KO and control cells between days 5-15 (E) or with expression of exogenous Flag-STN1 on day 15 (F). n=≥3 independent, biological replicates. Average values in the graphs indicate the mean and error bars denote ±s.e.m. P -values were calculated by a two-tailed, unpaired t -test. DOX=Doxycycline. D=day after DOX addition.

Article Snippet: Primary: OBFC1 (STN1) (Novus, NBP2-01006), Actinin (Santa Cruz, sc17829), pH3 S10 (Cell Signaling, 9706), RPA32 (Abcam, ab16850), and γH2AX (Bethyl, A300-081A; Abcam, ab81299).

Techniques: Control, Expressing, Two Tailed Test

Journal: bioRxiv

Article Title: Conditional deletion of human STN1 leads to telomere dysfunction and telomerase-dependent genome instability and proliferation defects

doi: 10.1101/2025.08.06.669015

Figure Lengend Snippet: Cells were treated with a telomerase inhibitor (BIBR) or DMSO starting with the addition of DOX on day 0 to the sgSTN1 cells. The control cells were also treated with BIBR or DMSO across the same timeframe. (A-B) Percentage of nuclei with 3 or more RPA (A) or RPA-γH2AX (B) foci in STN1 KO or control cells with or without BIBR treatment. n=3 independent, biological replicates. (C-D) Percentage of micronuclei (C) or anaphase bridges (D) on day 15 after DOX addition with and without BIBR treatment. n=4 independent, biological replicates. (E) Growth curve analysis, as indicated. n=3 independent, biological replicates. Average values in the graphs indicate the mean and error bars denote ±s.e.m. P -values were calculated by a two-tailed, unpaired t -test. DOX=Doxycycline. D=day after DOX addition.

Article Snippet: Primary: OBFC1 (STN1) (Novus, NBP2-01006), Actinin (Santa Cruz, sc17829), pH3 S10 (Cell Signaling, 9706), RPA32 (Abcam, ab16850), and γH2AX (Bethyl, A300-081A; Abcam, ab81299).

Techniques: Control, Two Tailed Test