Journal: Investigative ophthalmology & visual science

Article Title: Protective Effect of Met12, a Small Peptide Inhibitor of Fas, on the Retinal Pigment Epithelium and Photoreceptor After Sodium Iodate Injury.

doi: 10.1167/iovs.16-21392

Figure Lengend Snippet: FIGURE 1. Systemic administration of NaIO3 results in Fas activation in the retina and RPE. (A, B) There was a significant increase in the level of Fas receptor and caspase 3 transcript at 1 and 3 days post exposure to NaIO3. (C) The increased Fas-receptor transcript resulted in increased Fas receptor in the middle and outer retinal layers, as seen on immunohistochemistry. (D, E) There was an increased level of FasL in the RPE at both the transcript and protein levels, as detected by RT-PCR and immunohistochemistry, respectively. Of note, the isotype control did not show any nonspecific staining. *P < 0.05, **P < 0.01. Scale bars: 20 lm. INL, inner nuclear layer; ONL, outer nuclear layer; IS, inner segment; OS, outer segment; RPE65, retinal pigment epithelium protein of 65-kD molecular weight.

Article Snippet: Primary antibodies were as follows: Fas, 1:100 (Santa Cruz, Dallas, TX, USA); FasL, 1:100 (ab15285; Abcam, Cambridge, MA, USA); RPE65, 1:200 (a kind gift from Debra Thompson, University of Michigan, Ann Arbor, Michigan, USA); high mobility group box 1 (HMGB1), 1:200 (NB100-2322S; Novus Biologicals).

Techniques: Activation Assay, Immunohistochemistry, Reverse Transcription Polymerase Chain Reaction, Control, Staining, Molecular Weight

Journal: Scientific reports

Article Title: Graft cell expansion from hiPSC-RPE strip after transplantation in primate eyes with or without RPE damage.

doi: 10.1038/s41598-024-60895-w

Figure Lengend Snippet: Figure 2. Temporal changes in the RPE layer following micropulse laser treatment. (A) Retinal photographs of the eyes 2 and 5 days after the application of the subthreshold micropulse laser. (B) Autofluorescence images 2 and 5 days after the subthreshold microsecond pulse laser treatment. (C) Optical coherence tomography (OCT) images of the coagulated site 2 and 5 days after the microsecond pulse laser treatment. The yellow line indicates the locus of laser treatment. (D) Hematoxylin and eosin staining of the retinal pigment epithelium (RPE) damaged site immediately after and 2 and 5 days after the microsecond pulse laser treatment. (E) Immunostaining of RPE65 and rhodopsin at intact and RPE-damaged areas immediately after and 2 days after the microsecond pulse laser treatment. Yellow line indicates the locus of laser treatment. ONL outer nuclear layer. Scale bars: (D) 100 μm, (E) 500 μm.

Article Snippet: For the co-immunostaining using two different mouse antibodies for Ret P1 and RPE65, the FlexAble CoraLite®488 antibody labeling kit for mouse IgG1 (Proteintech KFA021) was used according to the manufacturer’s instructions.

Techniques: Tomography, Staining, Immunostaining

Journal: Scientific reports

Article Title: Graft cell expansion from hiPSC-RPE strip after transplantation in primate eyes with or without RPE damage.

doi: 10.1038/s41598-024-60895-w

Figure Lengend Snippet: Figure 4. Histology of hiPSC-RPE strip transplant at an RPE-damaged site (M4). (A) Low magnified hematoxylin and eosin (HE) image and the immunostaining image of the neighboring section with anti-human MERTK (hMERTK) and − RPE65 antibody in the hiPSC-RPE strip transplanted area over RPE damage. Yellow line indicates MERTK positive grafted area. (B) Immunostaining image of hMERTK and rhodopsin. Yellow line indicates hMERTK positive grafted area. (C) Engrafted RPE cells are positive for STEM121 and hMERTK on the apical side (arrows) while host RPE cells at the border show only autofluorescence but not STEM121 or hMERTK positivity (C’). (D) hMERTK on the apical side of transplanted RPE cells (arrows) face and interact with rhodopsin-positive outer segments of host photoreceptors. (E) Immunohistochemistry of type IV collagen in intact and RPE-damaged sites at 2 days after laser treatment. Expression of type IV collagen is disrupted after microsecond pulse laser treatment. (F) Expression analysis of type IV collagen in the intact, laser-damaged, and transplant sites. Expression of type IV collagen is identified at the basement membrane of transplanted RPE. (G) Expression analysis of type IV collagen at the basement membrane of hMERTK-positive RPE cells. Magnified view of yellow boxed part on the right. ONL outer nuclear layer, INL inner nuclear layer. Scale bars: (A) 200 μm, (B) 100 μm, (C,D) 10 μm, (E) 50 μm, (F) 100 μm, (G) 50 μm.

Article Snippet: For the co-immunostaining using two different mouse antibodies for Ret P1 and RPE65, the FlexAble CoraLite®488 antibody labeling kit for mouse IgG1 (Proteintech KFA021) was used according to the manufacturer’s instructions.

Techniques: Stripping Membranes, Immunostaining, Immunohistochemistry, Expressing, Membrane

Journal: Stem cell research

Article Title: Generation of a homozygous LRP2 knockout human embryonic stem cell line (FDCHDPe010-A-56) by CRISPR/Cas9 system

doi: 10.1016/j.scr.2021.102342

Figure Lengend Snippet: Reagents details.

Article Snippet: , Mouse anti-RPE65 , 1:100 , Novus Biologicals Cat# NB100-35 RRID:.

Techniques: Immunocytochemistry, Immunostaining, Flow Cytometry, Western Blot, Staining, Selection, Sequencing, Mutagenesis