rbpms polyclonal antibody  (Proteintech)

Journal: Cell Death & Disease

Article Title: Reln-Dab1 pathway mitigates retinal ganglion cell apoptosis in retinal ischemia-reperfusion injury

doi: 10.1038/s41419-025-07742-6

Figure Lengend Snippet: A Schematic representation of the experimental design (Created with BioRender.com). B Histopathological examination of retinal tissue stained with H&E showed that the inner retinal thickness of mice was significantly reduced after I/R injury. This I/R-induced inner retinal thinning was significantly improved in mice treated with exogenous Reelin protein. Scale bar: 80 μm. C Quantification of inner retinal thickness ( n = 6). D Representative OCT images of retinal tissue demonstrate changes after I/R injury. E Quantification of total retinal thickness ( n = 6). F Representative fluorescent staining of retinal patches after I/R injury showed the RBPMS-positive RGCs (orange) distribution. Supplementation with Reelin protein exerted a significant protective effect on I/R-induced RGC injury. Scale bars: 100 μm or 1 mm. G Number of RBPMS-positive RGCs ( n = 4). H Representative oscillatory potential waveforms of visual function were assessed via FERG 7 days after I/R injury. I Quantification of the magnitude of a and b waves ( n = 6). Data are represented as means ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: After removing pigmented tissue and rinsing in 0.1% PBST, retinas were dehydrated in methanol, blocked with goat serum, and incubated with the RGC marker RBPMS polyclonal antibody (Proteintech, Wuhan, China) overnight at 4 °C.

Techniques: Staining

Journal: Cell Death & Disease

Article Title: Reln-Dab1 pathway mitigates retinal ganglion cell apoptosis in retinal ischemia-reperfusion injury

doi: 10.1038/s41419-025-07742-6

Figure Lengend Snippet: A Schematic representation of the experimental design (Created with BioRender.com). B Frozen slides showing retinal infection 3 weeks after intravitreal injection of AAV. EGFP carried by AAV was expressed in nearly all the layers of the retina. Scale bar: 40 μm. C The expression level of Reelin protein in the inner retinal layer was assessed by the intensity of the fluorescence signals ( n = 6). D The mRNA expression of the Reln gene ( n = 6). E Histopathological examination of retinal tissue via H&E staining showed that the inner retinal thickness in AAV- shReln mice was further reduced after I/R injury. The damage to I/R-induced RGCs was further aggravated in AAV- shReln mice. Scale bar: 80 μm. F Quantification of inner retinal thickness ( n = 6). G Representative OCT images of retinal tissue changes after I/R injury. H Quantification of total retinal thickness ( n = 6). I Representative fluorescent staining of retinal patches after I/R injury showed the distribution of RBPMS-positive RGCs (orange). Scale bars: 100 μm or 1 mm. J Number of RBPMS-positive RGCs ( n = 4). K Representative oscillatory potential waveforms of visual function were assessed by FERG after I/R injury. L Quantification of the magnitude of a and b waves ( n = 6). Data are represented as means ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: After removing pigmented tissue and rinsing in 0.1% PBST, retinas were dehydrated in methanol, blocked with goat serum, and incubated with the RGC marker RBPMS polyclonal antibody (Proteintech, Wuhan, China) overnight at 4 °C.

Techniques: Infection, Injection, Expressing, Fluorescence, Staining

Journal: Stem Cell Research & Therapy

Article Title: Retinal ganglion cells induce stem cell-derived neuroprotection via IL-12 to SCGF-β crosstalk

doi: 10.1186/s13287-025-04198-5

Figure Lengend Snippet: Intravitreal injection of SCGF-β protects RGCs after optic nerve crush. A Human SCGF-β or PBS was intravitreally injected into the adult mouse eye right after optic nerve crush. The retina was explanted and immunostained 5 days post cush. B RBPMS + RGCs under SCGF-β administration showed significantly higher survival, compared with the PBS treatment. * P < 0.05. ONC, optic nerve crush. Error bar denotes SD

Article Snippet: The flat mounted samples were permeabilized with 0.3% Triton X-100 (Cat. #T9284; Sigma-Aldrich) for 20 min, blocked with 5% normal goat serum (Cat. #16,210,064; Invitrogen, San Diego, CA, USA) in PBS for 1 h. For the retinas from optic nerve crush, the samples were incubated with a rabbit polyclonal anti-RNA Binding Protein, mRNA Processing Factor (RBPMS) antibody (1:500; Cat. #1832-RBPMS; PhosphoSolutions, Aurora, CO, USA) overnight at 4 °C, rinsed three times with PBS, and then incubated with Alexa Fluor 555-tagged secondary antibody (1:500; Cat. #A-21428; Life Technologies) overnight, again.

Techniques: Injection

Journal: Stem Cell Research & Therapy

Article Title: Retinal ganglion cells induce stem cell-derived neuroprotection via IL-12 to SCGF-β crosstalk

doi: 10.1186/s13287-025-04198-5

Figure Lengend Snippet: RGCs enhance iPSC’s SCGF-β release via IL-12(p70). A - B IL-12(p70) was detected in RGC supernatant, not in RGC medium, through multiplexed antibody-based assays and confirmed by ELISA. C - D Compared to the control (“No IL-12(p70)” group), significant upregulation of SCGF-β (via ELISA ) and CLEC11A mRNA (via qRT-PCR ) were observed in iPSCs treated with 2.5, 5, and 10 ng/mL of IL-12(p70). The expression peaked in the 5 ng/mL IL-12(p70)-treated group. E RGCs were cultured in the RGC medium with different dosages of IL-12(p70) administration. A significantly greater cell viability was not found until treated with 40 ng/mL of IL-12(p70). * P < 0.05. Error bar denotes SD. NS, nonsignificant

Article Snippet: The flat mounted samples were permeabilized with 0.3% Triton X-100 (Cat. #T9284; Sigma-Aldrich) for 20 min, blocked with 5% normal goat serum (Cat. #16,210,064; Invitrogen, San Diego, CA, USA) in PBS for 1 h. For the retinas from optic nerve crush, the samples were incubated with a rabbit polyclonal anti-RNA Binding Protein, mRNA Processing Factor (RBPMS) antibody (1:500; Cat. #1832-RBPMS; PhosphoSolutions, Aurora, CO, USA) overnight at 4 °C, rinsed three times with PBS, and then incubated with Alexa Fluor 555-tagged secondary antibody (1:500; Cat. #A-21428; Life Technologies) overnight, again.

Techniques: Enzyme-linked Immunosorbent Assay, Control, Quantitative RT-PCR, Expressing, Cell Culture

Journal: Stem Cell Research & Therapy

Article Title: Retinal ganglion cells induce stem cell-derived neuroprotection via IL-12 to SCGF-β crosstalk

doi: 10.1186/s13287-025-04198-5

Figure Lengend Snippet: iPSC-derived SCGF-β promotes RGC survival via upregulation of ngn2. A - B RT-qPCR results showed a significant increase of ngn2 mRNA in RGCs cocultured with iPSCs or treated with SCGF-β. C - D Overexpression of ngn2 in RGCs cultured in the RGC medium for 1 week significantly increased RGC viability. E Knockdown of ngn2 in RGCs cultured for 1 week does not significantly alter RGC survival, whether treated with SCGF-β or not. * P < 0.05. Error bar denotes SD

Article Snippet: The flat mounted samples were permeabilized with 0.3% Triton X-100 (Cat. #T9284; Sigma-Aldrich) for 20 min, blocked with 5% normal goat serum (Cat. #16,210,064; Invitrogen, San Diego, CA, USA) in PBS for 1 h. For the retinas from optic nerve crush, the samples were incubated with a rabbit polyclonal anti-RNA Binding Protein, mRNA Processing Factor (RBPMS) antibody (1:500; Cat. #1832-RBPMS; PhosphoSolutions, Aurora, CO, USA) overnight at 4 °C, rinsed three times with PBS, and then incubated with Alexa Fluor 555-tagged secondary antibody (1:500; Cat. #A-21428; Life Technologies) overnight, again.

Techniques: Derivative Assay, Quantitative RT-PCR, Over Expression, Cell Culture, Knockdown

Journal: Stem Cell Research & Therapy

Article Title: Retinal ganglion cells induce stem cell-derived neuroprotection via IL-12 to SCGF-β crosstalk

doi: 10.1186/s13287-025-04198-5

Figure Lengend Snippet: Overexpression of ngn2 protects endogenous and transplanted RGCs in vivo. A AAV2-ngn2-EGFP or AAV2-EGFP particles were intravitreally injected 2 weeks into adult mouse eyes 2 weeks before optic nerve crush. Retina explants were harvested and immunostained 2 weeks after the optic nerve crush. Surviving RGCs were labeled with RBPMS (red). B A significantly greater RBPMS + RGC number was found on retina explants from the AAV-ngn2 treated group. C The EGFP and mCherry were used to label the entire transplanted and ngn2-overexpressing donor mouse RGCs, respectively. Ngn2-mCherry-overexpressing mouse RGCs that were transplanted into adult rat eyes shows significantly higher survival rates, compared with the negative control RGC transplant 1 week after transplantation in vivo. D & E Difference between transplanted RGC survival rates and average neurite lengths with and without ngn2 overespression was quantified . Donor RGCs overexpressing ngn2-mCherry grew significantly longer neurites than those from negative control-RGC transplantation in vivo. * P < 0.05, paired t-test. ONC, optic nerve crush; NC, negative control; OE, overexpression. Error bar denotes SD

Article Snippet: The flat mounted samples were permeabilized with 0.3% Triton X-100 (Cat. #T9284; Sigma-Aldrich) for 20 min, blocked with 5% normal goat serum (Cat. #16,210,064; Invitrogen, San Diego, CA, USA) in PBS for 1 h. For the retinas from optic nerve crush, the samples were incubated with a rabbit polyclonal anti-RNA Binding Protein, mRNA Processing Factor (RBPMS) antibody (1:500; Cat. #1832-RBPMS; PhosphoSolutions, Aurora, CO, USA) overnight at 4 °C, rinsed three times with PBS, and then incubated with Alexa Fluor 555-tagged secondary antibody (1:500; Cat. #A-21428; Life Technologies) overnight, again.

Techniques: Over Expression, In Vivo, Injection, Labeling, Negative Control, Transplantation Assay