mrna processing factor rbpms  (Proteintech)

Journal: Neural regeneration research

Article Title: Interleukin-4 promotes microglial polarization toward a neuroprotective phenotype after retinal ischemia/reperfusion injury.

doi: 10.4103/1673-5374.339500

Figure Lengend Snippet: Figure 1 ｜RGCs are lost continuously over a 28-day period after RIR injury. (A) Schematic diagram of the strategy used to induce RIR injury. A 30-gauge needle connected to 1.2-meter tall saline container was cannulated in the anterior chamber of the mouse eye. (B) Illustration of the strategy used for counting cells in retinal flat mounts. The image on the left shows a whole retinal flat mount photographed at 100× magnification (red: IBA1 [ionized calcium binding adaptor molecule 1], scale bar: 1 mm), and the image on the right shows a part of the flat mount photographed at 200× magnification (scale bar: 50 μm) for cell counting. Each retina was divided into four quadrants with 3 different regions each (highlighted by 12 small white boxes in the retina), i.e., the central, intermediate, and peripheral regions, and the images were captured for cell counting as shown in the right panel. The average cell numbers in these 12 regions were used to calculate the cell density for the entire retina. Changes in RGC numbers on days 0, 1, 3, 7, 14, and 28 after RIR injury are shown in (C) and (D). RNA binding protein, mRNA processing factor (RBPMS; (red) was used as a marker for RGCs. (C) Representative images for the assessment of RBPMS expression within the whole retina (scale bar: 50 µm) showed that RGC loss occurred in a homogeneous fashion over the entire retina, including the region close to the optic nerve head and in the periphery, and the loss became gradually more severe over time. (D) Representative images for RBPMS+ cell counting (scale bar: 1 mm). The RBPMS+ cell numbers decreased and the RBPMS– regions expanded gradually until almost the entire retina was RBPMS– by day 28 after RIR surgery. (E) The quantification of RBPMS+ cells in panel D, which represents the average RGC number in retinas from mice at 0, 1, 3, 7, 14, and 28 days after RIR. The bar chart indicated that RGCs were lost continuously within the 28-day period following RIR. Among adjacent time points, the largest RGC reduction occurred between days 1 and 3. On days 7 and 28, the number of RGCs decreased to approximately 50% and 25% of that observed on day 0, respectively. n = 4 mice per group. Data are expressed as the means ± SD; *P < 0.05, ***P < 0.001 (unpaired Student’s t-tests). Three independent replicates were measured in each experiment. RGC: Retinal ganglion cell; RIR: retinal ischemia/ reperfusion.

Article Snippet: The following primary antibodies were used: rabbit anti-ionized calcium binding adaptor molecule 1 (IBA1; marker for retinal microglia; 1:1000, Cat# 019-19741, Wako Pure Chemical Industries, Osaka, Japan), goat anti-CD206 (marker for M2 microglia; 1:800, Cat# AF2535, R&D Systems, Minneapolis, MN, USA), rabbit anti-RNA binding protein, mRNA processing factor (RBPMS; marker for RGCs; 1:200, Cat# NBP2-20112, Novus Biologicals, Manchester, UK), and goat anti-ARG1 (1:100, Cat# ab60176, Abcam, Cambridge, UK).

Techniques: Saline, Binding Assay, Cell Counting, RNA Binding Assay, Marker, Expressing

Journal: Neural regeneration research

Article Title: Interleukin-4 promotes microglial polarization toward a neuroprotective phenotype after retinal ischemia/reperfusion injury.

doi: 10.4103/1673-5374.339500

Figure Lengend Snippet: Figure 4 ｜Flowchart of Experiment II. IL: Interleukin; PBS: phosphate-buffered saline; qRT-PCR: quantitative reverse transcription-polymerase chain reaction; RBPMS: RNA binding protein, mRNA processing factor; RGC: retinal ganglion cell.

Article Snippet: The following primary antibodies were used: rabbit anti-ionized calcium binding adaptor molecule 1 (IBA1; marker for retinal microglia; 1:1000, Cat# 019-19741, Wako Pure Chemical Industries, Osaka, Japan), goat anti-CD206 (marker for M2 microglia; 1:800, Cat# AF2535, R&D Systems, Minneapolis, MN, USA), rabbit anti-RNA binding protein, mRNA processing factor (RBPMS; marker for RGCs; 1:200, Cat# NBP2-20112, Novus Biologicals, Manchester, UK), and goat anti-ARG1 (1:100, Cat# ab60176, Abcam, Cambridge, UK).

Techniques: Saline, Quantitative RT-PCR, Reverse Transcription, Polymerase Chain Reaction, RNA Binding Assay

Journal: Neural regeneration research

Article Title: Interleukin-4 promotes microglial polarization toward a neuroprotective phenotype after retinal ischemia/reperfusion injury.

doi: 10.4103/1673-5374.339500

Figure Lengend Snippet: Figure 7 ｜Administration of IL-4 prolonged the duration of M2 microglia polarization and improved RGC survival. (A) Representative images of retinal flat mounts stained for IBA1 (red) and ARG1 (blue) after treatment with IL-4 or vehicle. These photos suggested that the densities of ARG1+ cells were elevated markedly in RIR mice after IL-4 administration. Scale bar: 50 µm. All fluorescent images in this study were captured by laser confocal microscopy. In contrast to conventional fluorescence microscopy, the colors of the images were pseudo-colors selected by the user rather than the true dye color. We chose blue as the color of ARG1 out of personal preference. (B) Quantification of the percentage of ARG1+ areas in injured retinas treated with IL-4 or vehicle. Retinas from the IL-4 group displayed a greater than 2-fold increase in the ARG1+ area (%) compared with those in the vehicle group (n = 4 mice per group). (C) The mRNA expression of IL4rα and M2-related genes in retinas from the vehicle and IL-4 treatment groups. IL-4 treatment led to a 6–7-fold increase in both Arg1 and Cd206 mRNA and approximately 1.8-fold increase in IL4rα mRNA compared with vehicle (n = 3–5 per group). (D) Representative images of RBPMS expression (red) in retinas from mice 7 days after the last intravitreal dose of IL-4. The upper (scale bar: 50 µm) and lower (scale bar: 1 mm) images were used for cell counting and overall evaluation of RBPMS expression within the whole retina, respectively. The density of RBPMS+ cells in the IL-4 group was higher than that in the vehicle group but less than that in the normal group (RIR and IL-4 negative). (E) Quantification of RGC numbers in retinas from normal, vehicle, and IL-4 groups. The RGC numbers in the IL-4 group were approximately twice greater than that in the vehicle group (n = 4–5 mice per group). Data are presented as means ± SD. *P < 0.05, **P < 0.01, vs. vehicle (unpaired Student’s t-tests). Three independent replicates were measured for each experiment. IBA1: Ionized calcium binding adaptor molecule 1; IL: interleukin; RBPMS: RNA binding protein, mRNA processing factor; RGC: retinal ganglion cell; RIR: retinal ischemia reperfusion

Article Snippet: The following primary antibodies were used: rabbit anti-ionized calcium binding adaptor molecule 1 (IBA1; marker for retinal microglia; 1:1000, Cat# 019-19741, Wako Pure Chemical Industries, Osaka, Japan), goat anti-CD206 (marker for M2 microglia; 1:800, Cat# AF2535, R&D Systems, Minneapolis, MN, USA), rabbit anti-RNA binding protein, mRNA processing factor (RBPMS; marker for RGCs; 1:200, Cat# NBP2-20112, Novus Biologicals, Manchester, UK), and goat anti-ARG1 (1:100, Cat# ab60176, Abcam, Cambridge, UK).

Techniques: Staining, Confocal Microscopy, Fluorescence, Microscopy, Expressing, Cell Counting, Binding Assay, RNA Binding Assay

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

Journal: Pharmaceutics

Article Title: Retinal Protection of New Nutraceutical Formulation

doi: 10.3390/pharmaceutics17010073

Figure Lengend Snippet: Immunostaining of RNA-binding protein and mRNA Processing Factor-positive (RBPMS+) cells for the analysis of retinal ganglion cell (RGC) density in flat-mount retinas. ( a ) RBPMS+ RGCs in central ( A , B ) and peripheral areas ( C , D ) of control (CTRL) and Epicolin formulation-treated mice. Quantification was carried out on the whole retina ( b ) and on both central and peripheral areas of the retina ( c ). Scale bar corresponds to 50 μm. The data were plotted as means ± SEMs (n = 12 retinas per group). * p ≤ 0.05 vs. CTRL; ° p ≤ 0.05 vs. CTRL c; † p ≤ 0.05 vs. CTRL p. Unpaired t -test. CTRL central (CTRL c), CTRL periphery (CTRL p), Epicolin central (Epicolin c), and Epicolin periphery (Epicolin p).

Article Snippet: Primary antibody for RNA binding protein, mRNA Processing Factor (RBPMS, RNA-binding protein with multiple splicing), was purchased from Novus Biologicals, part of Bio-Techne SRL (Milan, Italy).

Techniques: Immunostaining, RNA Binding Assay, Control, Formulation