========================
Location in DDF requires
========================

not Brca1--

H2ax-- phos site S140

  *** Rnf8[Ab] locatedin(subnuclear-bodies)[IHC] is detectable
    *** cells: mEFs<H2ax~null><xH2ax> in BMS "treated with IR for 6 hr"
    *** inhibited by: xH2ax(S140A) [substitution]
    *** source: 18001825-Fig-S1d
  *** Rnf8[tAb] locatedin(subnuclear-bodies)[IHC] is detectable
    *** cells: mEFs<xRnf8> in BMS "treated with IR for 6 hr"
    *** reqs: H2ax [KO]
    *** source: 18001825-Fig-1e

Mdc1--  phos sites in TQXF motif (T699/T719/T752/T765)

  -------------- reqs phos sites in TQXF motif (T699/T719/T752/T765) ---
  *** xRnf8[XFP] locatedin(subnuclear-bodies)[IHC] is detectable
    *** cells: U2OS<Mdc1~kd><xMdc1><xRnf8> in BMS "treated with IR for 30 min"
    *** inhibited by: xMdc1(T699A/T719A/T752A) [substitution]
    *** source: 18001824-Fig-2g
  *** Rnf8[Ab] locatedin(subnuclear-bodies)[IHC] is detectable
    *** cells: HELA<Mdc1~kd><xMdc1> in BMS "treated with IR for 6 hr"
    *** reqs: xMdc1 [omission]
    *** inhibited by: xMdc1(del(698-768)) [substitution]
    *** source: 18001825-Fig-S3b
  -------------------------------------------------------------------
  *** xRnf8[XFP] locatedin(subnuclear-bodies)[IHC] is detectable
    *** cells: U2OS<xRnf8> in BMS "treated with IR for 30 min"
    *** reqs: Mdc1 [RNAi]
    *** source: 18001824-Fig-2g
  *** Rnf8[tAb] locatedin(subnuclear-bodies)[IHC] is detectable
    *** cells: mEFs<xRnf8> in BMS "treated with IR for 6 hr"
    *** reqs: Mdc1 [KO]
    *** source: 18001825-Fig-1e
  *** xRnf8[XFP] locatedin(DSBs)[IHC] is detectable
    *** cells: U2OS<xRnf8> in BMS "treated with MicroLaser for 30 min"
    *** reqs: Mdc1 [RNAi]
    *** source: 18001824-Fig-1f

not Nbs1--

  *** Rnf8[tAb] locatedin(subnuclear-bodies)[IHC] is detectable
    *** cells: NotReported<Nbs1~fmut><xNbs1> in BMS "treated with IR for 6 hr"
    *** comment: colocates with H2ax{phos(S140)}[phosAb]
    *** does not req: xNbs1 [omission]
    *** unaffected by: xNbs1(S343A) [substitution]
    *** source: 18001825-Fig-1e

not Tp53bp1--

  *** Rnf8[tAb] locatedin(subnuclear-bodies)[IHC] is detectable
    *** cells: mEFs<xRnf8> in BMS "treated with IR for 6 hr"
    *** comment: colocates with H2ax{phos(S140)}[phosAb]
    *** does not req: Tp53bp1 [KO]
    *** source: 18001825-Fig-1e

not Ubc13--

  *** Rnf8[Ab] locatedin(subnuclear-bodies)[IHC] is detectable
    *** cells: HELA in BMS "treated with IR for 6 hr"
    *** does not req: Ubc13 [RNAi]
    *** source: 18001825-Fig-S5d

Rnf8 features  FHA domain not Ring domain

  *** xRnf8[tAb] locatedin(subnuclear-bodies)[IHC] is detectable
    *** cells: HELA<Rnf8~kd><xRnf8> in BMS "treated with IR for 6 hr"
    *** inhibited by: xRnf8(mnr)"delFHA" [substitution]
    *** unaffected by: xRnf8(mnr)"delRING" [substitution]
    *** source: 18001825-Fig-4c
  *** xRnf8[tAb] locatedin(subnuclear-bodies)[IHC] is detectable
    *** cells: HELA<Rnf8~kd><xRnf8> in BMS "treated with IR for 6 hr"
    *** inhibited by: xRnf8(mnr)"delFHA" [substitution]
    *** unaffected by: xRnf8(mnr)"delRING" [substitution]
    *** source: 18001825-Fig-5c
  *** xRnf8[tAb] locatedin(subnuclear-bodies)[IHC] is detectable
    *** cells: HELA<Rnf8~kd><xRnf8> in BMS "treated with IR for 6 hr"
    *** inhibited by: xRnf8(mnr)"delFHA" [substitution]
    *** unaffected by: xRnf8(mnr)"delRING" [substitution]
    *** source: 18001825-Fig-5d
  *** xRnf8[tAb] locatedin(subnuclear-bodies)[IHC] is detectable
    *** cells: HELA<Rnf8~kd><xRnf8> in BMS "treated with IR for 6 hr"
    *** inhibited by: xRnf8(mnr)"delFHA" [substitution]
    *** unaffected by: xRnf8(mnr)"delRING" [substitution]
    *** source: 18001825-Fig-S4f
  *** xRnf8[XFP] locatedin(subnuclear-bodies)[IHC] is increased irt IR (30 min)
    *** cells: U2OS<xRnf8> in BMS
    *** inhibited by: xRnf8(R42A)"mutFHA" [substitution]
    *** unaffected by: xRnf8(C403S)"mutRING" [substitution]
    *** source: 18001824-Fig-2c
  *** xRnf8[tAb] locatedin(subnuclear-bodies)[IHC] is increased irt IR (6 hr)
    *** cells: HEK293T<xRnf8> in BMS
    *** comment: colocates with H2ax{phos(S140)}[phosAb]
    *** inhibited by: xRnf8(mnr)"delFHA" [substitution]
    *** unaffected by: xRnf8(mnr)"delRING" [substitution]
    *** source: 18001825-Fig-1f


chemicals

  *** xRnf8[XFP] locatedin(DSBs)[IHC] is detectable
    *** cells: U2OS<xRnf8> in BMS "treated with MG132 for 90 min followed by MicroLaser for 30 min"
    *** source: 18001824-Fig-3c

no extras

  *** Rnf8[Ab] locatedin(subnuclear-bodies)[IHC] is detectable
    *** cells: NotReported in BMS "treated with IR for 6 hr"
    *** source: 18001825-Fig-S1c
  *** xRnf8[tAb] locatedin(subnuclear-bodies)[IHC] is detectable
    *** cells: HEK293T<xRnf8> in BMS "treated with IR for 6 hr"
    *** source: 18001825-Fig-1a
  *** Rnf8[Ab] locatedin(subnuclear-bodies)[IHC] is detectable
    *** cells: HELA in BMS "treated with IR for 6 hr"
    *** source: 18001825-Fig-4a
  *** Rnf8[Ab] locatedin(DSBs)[IHC] is detectable
    *** cells: U2OS in BMS "treated with MicroLaser for 2 min"
    *** source: 18001824-Fig-S4b
  *** xRnf8[XFP] locatedin(DSBs)[IHC] is detectable
    *** cells: U2OS<xRnf8> in BMS "treated with MicroLaser for 30 min"
    *** source: 18001824-Fig-1c
  *** xRnf8[XFP] locatedin(DSBs)[IHC] is increased irt MicroLaser (14 min)
    *** cells: U2OS<xRnf8> in BMS
    *** comment: increase was monitored continuously during MicroLaser
    *** comment: increase was similar to xNbs1 xMdc1 in same system
    *** comment: increase was faster than xBrca1 xTp53bp1 xAtr in same system
    *** source: 18001824-Fig-3b
  *** xRnf8[XFP] locatedin(DSBs)[IHC] is increased irt MicroLaser (times)
    *** cells: U2OS<xRnf8> in BMS
    *** times: 0 30+ 60++ 120+++ 360++++ sec
    *** source: 18001824-Fig-3a
  *** Rnf8[Ab] locatedin(subnuclear-bodies)[IHC] is increased irt IR (6 hr)
    *** cells: HEK293T in BMS
    *** source: 18001825-Fig-1b
  *** xRnf8[XFP] locatedin(subnuclear-bodies)[IHC] is increased irt IR (30 min)
    *** cells: U2OS<xRnf8> in BMS
    *** source: 18001824-Fig-1b
  *** xRnf8[XFP] locatedin(subnuclear-bodies)[IHC] is increased irt IR (times)
    *** cells: U2OS<xRnf8> in BMS
    *** times: 0 1++ 2++ 4++ 6++ 8+ 24 hr
    *** source: 18001824-Fig-S4c
