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. 2017 Jun 12;18(8):1367–1381. doi: 10.15252/embr.201643535

Figure 4. Co‐localizations and interactions of BAM3, CRN, and CLV2.

Figure 4

  • A
    Transient expression of CRN‐TurboRFP fusion protein (red fluorescence) in tobacco (Nicotiana benthamiana) leaf epidermal cells, under control of a constitutive promoter (confocal microscopy).
  • B
    Transient expression of CLV2‐CITRINE fusion protein (green fluorescence) in tobacco leaf epidermal cells.
  • C
    Transient co‐expression of CRN‐TurboRFP fusion protein and (non‐fluorescent) CLV2‐HA fusion protein in tobacco leaf epidermal cells.
  • D
    Transient co‐expression of CRN‐TurboRFP and CLV2‐CITRINE fusion proteins in tobacco leaf epidermal cells. Red fluorescent and green fluorescent channels are shown separately and in overlay.
  • E, F
    Expression pattern of CRN‐CITRINE fusion protein (green fluorescence) under control of the BAM3 promoter in crn (E) or clv2 mutant (F), with corresponding close‐ups (magenta fluorescence: calcofluor white cell wall staining). Asterisks mark developing protophloem sieve element strands. Arrowhead in (E′) highlights plasma membrane‐localized CRN‐CITRINE.
  • G
    Transient co‐expression of CRN‐TurboRFP and BAM3‐CITRINE fusion proteins in tobacco leaf epidermal cells. Red fluorescent and green fluorescent channels are shown separately and in overlay.
  • H
    Transient co‐expression of CRN‐TurboRFP, BAM3‐CITRINE, and (non‐fluorescent) CLV2‐HA fusion proteins in tobacco leaf epidermal cells. Red fluorescent and green fluorescent channels are shown separately and in overlay.
  • I
    Bimolecular fluorescence complementation (BiFC) between transiently co‐expressed CLV2 and CRN proteins fused to one half each of YFP (yellow fluorescence) in the presence of co‐expressed BAM3‐mTFP1 fusion protein (blue fluorescence). Yellow fluorescent and blue fluorescent channels are shown separately and in overlay.
  • J
    BiFC between transiently co‐expressed BAM3 and CRN proteins fused to one half each of YFP (green fluorescence) in the presence of co‐expressed (non‐fluorescent) CLV2‐HA fusion protein in tobacco leaf epidermal cells (J′′). Parallel control experiments for BiFC between CLV2 and CRN (J) and BAM3 and CRN (J′) are shown.
  • K
    BiFC between transiently co‐expressed BAM3 and MAKR5 proteins fused to one half each of YFP (red: chloroplast autofluorescence).
  • L
    BiFC between transiently co‐expressed BRI1 and BKI1 proteins fused to one half each of YFP (green fluorescence).
  • M
    BiFC between transiently co‐expressed BRI1 and CRN proteins fused to one half each of YFP in the presence of co‐expressed (non‐fluorescent) CLV2‐HA fusion protein in tobacco leaf epidermal cells (red: chloroplast autofluorescence).
  • N, O
    Expression of BAM3‐CITRINE or CRN‐CITRINE fusion proteins (green fluorescence) under control of the BAM3 promoter in the developing sieve element cells close to the stem cells in Col‐0 (N) or crn (O) background (magenta fluorescence: calcofluor white cell wall staining). Red arrowhead indicates rootward direction.