DNA oligonucleotides used in Figure 5 are:
5’-TGGCGACGGCAGCGAGGCTTTTTTTTTTTTTTTTTTTT-iCy3-TTTTTTTTTTTT-3’-Cy5 5’-GCCTCGCTGCCGTCGCCA-3’-biotin
DNA oligonucleotides used inFigure S6 are:
5’-TGGCGACGGCAGCGAGGCTTTTTTTTTTTTTTTT-iCy3-TTTTTTTTTTTTTT-3’ Cy5–5’-GCCTCGCTGCCGTCGCCA-3’-biotin
For the inhibitors used in this study, cells were treated with inhibitors 3 hours before fixation. ATRi: VE-821 (Selleckchem), 2 μM, 3 hours incubation; CDC7i: PHA767491 (Selleckchem), 20μM, 3 hours incubation; Chk1i: UCN01 (Sigma-Aldrich), 100 nM, 1 hour incubation; Chk2i: PV1019 (Sigma-Aldrich), 5 μM, 3 hours incubation; For Hydroxyurea treatments (HU, Sigma-Aldrich), cells were treated 2 hours before fixation.
Antibodies against the imaged proteins are (ms = mouse; rb = rabbit; AF = AlexaFluor): ms-anti-PCNA (SCBT sc56, 1:1,000), rb-anti-MCM6 (AF568 conjugated, Abcam ab211916, 1:1,000), rb-anti-RPA1 (AF647 conjugated, Abcam ab199240, 1:10,000), rb-anti-pS4/S8RPA2 (Abcam ab87277, 1:1000). Secondary antibodies are: goat-anti-ms (AF488 conjugated, ThermoFisher, 1:10,000), goat-anti-rb (AF647 conjugated, ThermoFisher, 1:10,000).
Antibodies against the blotted proteins are:
Ms-anti-RPA2 (Millipore, MABE285, 1:1000), rb-anti-c-myc (Bethyl, A190–105A, 1:1000), ms-anti-β-tubulin (Sigma-Aldrich T8328, 1:1000), Ms-anti-HRP (Abcam, ab181708, 1: 10000), Rb-anti-HRP (Abcam, ab34885, 1:10000).
5’-TGGCGACGGCAGCGAGGCTTTTTTTTTTTTTTTTTTTT-iCy3-TTTTTTTTTTTT-3’-Cy5 5’-GCCTCGCTGCCGTCGCCA-3’-biotin
DNA oligonucleotides used in
5’-TGGCGACGGCAGCGAGGCTTTTTTTTTTTTTTTT-iCy3-TTTTTTTTTTTTTT-3’ Cy5–5’-GCCTCGCTGCCGTCGCCA-3’-biotin
For the inhibitors used in this study, cells were treated with inhibitors 3 hours before fixation. ATRi: VE-821 (Selleckchem), 2 μM, 3 hours incubation; CDC7i: PHA767491 (Selleckchem), 20μM, 3 hours incubation; Chk1i: UCN01 (Sigma-Aldrich), 100 nM, 1 hour incubation; Chk2i: PV1019 (Sigma-Aldrich), 5 μM, 3 hours incubation; For Hydroxyurea treatments (HU, Sigma-Aldrich), cells were treated 2 hours before fixation.
Antibodies against the imaged proteins are (ms = mouse; rb = rabbit; AF = AlexaFluor): ms-anti-PCNA (SCBT sc56, 1:1,000), rb-anti-MCM6 (AF568 conjugated, Abcam ab211916, 1:1,000), rb-anti-RPA1 (AF647 conjugated, Abcam ab199240, 1:10,000), rb-anti-pS4/S8RPA2 (Abcam ab87277, 1:1000). Secondary antibodies are: goat-anti-ms (AF488 conjugated, ThermoFisher, 1:10,000), goat-anti-rb (AF647 conjugated, ThermoFisher, 1:10,000).
Antibodies against the blotted proteins are:
Ms-anti-RPA2 (Millipore, MABE285, 1:1000), rb-anti-c-myc (Bethyl, A190–105A, 1:1000), ms-anti-β-tubulin (Sigma-Aldrich T8328, 1:1000), Ms-anti-HRP (Abcam, ab181708, 1: 10000), Rb-anti-HRP (Abcam, ab34885, 1:10000).