# kap gfp (Cell Signaling Technology Inc)

Cell Signaling Technology Inc is a verified supplier

Cell Signaling Technology Inc manufactures this product

## Structured Review

Kap Gfp, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more

https://www.bioz.com/result/kap gfp/product/Cell Signaling Technology Inc

Average 86 stars, based on 1 article reviews

Price from $9.99 to $1999.99

## Images

### 1) Product Images from "The ERK activator, BCI, inhibits ciliogenesis and causes defects in motor behavior, ciliary gating, and cytoskeletal rearrangement"

**Article Title: **The ERK activator, BCI, inhibits ciliogenesis and causes defects in motor behavior, ciliary gating, and cytoskeletal rearrangement

**Journal: **Life Science Alliance

**doi: **10.26508/lsa.202301899

**Figure Legend Snippet:**(A) Chlamydomonas expressing KAP-GFP. Indicated are the measured areas of KAP-GFP localization: the two basal bodies and cilia. Scale bar is 5 μm. (B) Super plot quantification of basal body fluorescence in (A) after a 2-h treatment with BCI at the indicated concentrations. Error bars are the mean with the 95% confidence interval for the averages from the trials. P = 0.5126, which was determined by an ordinary one-way ANOVA. (C) Ciliary length plotted against ciliary fluorescence for cells in DMSO (orange circles) or 30 μM BCI (purple) after a 2-h treatment (n = 40, N = 1). Simple linear regression was used to generate linear regression lines and comparisons. For DMSO, y = 0.1171x + 0.4305 and r 2 = 0.05676. For BCI, y = 0.1174x + 0.4051 and r 2 = 0.2636. Comparing slopes, F = 4.048e-006 (1, 76) and P = 0.9984. Comparing intercepts, F = 0.005429 (1, 77) and P = 0.9415. (D) Comparison of total ciliary fluorescence shown in (C). P -values were determined using a t test. (E) Comparisons of ciliary fluorescence in DMSO or BCI per micrometer of cilia in (C). P -values were determined using a t test. (F) Example kymographs collected from total internal reflection fluorescent microscopy of KAP-GFP movement in cilia in cells treated with DMSO (top) or 30 μM BCI (bottom) for 2 h. Vertical scale bars are 4 μm. Horizontal scale bars are 2 s. (G, H, I) KAP-GFP dynamics quantified from the kymographs (n = 20, N = 1). Error bars are the mean with the 95% confidence interval (n = 20, N = 1). P -values were calculated from a two-tailed unpaired t test for pairwise comparisons. (G) Frequency of KAP-GFP trains measured as the total amount of trains counted over the total amount of time the kymograph was collected. For DMSO, n = 40 (1 and 2 h). For BCI, n = 30 (1 h) and 34 (2 h). (H) Velocity of KAP-GFP trains measured as the distance traveled in μm over time in seconds. For DMSO, n = 100 (1 and 2 h). For BCI, n = 93 (1 h) and 88 (2 h). (I) Relative injection size of KAP-GFP trains measured as the relative total fluorescent intensity of each train relative to the maximum measurement. For DMSO, n = 100 (1 and 2 h). For BCI, n = 93 (1 h) and 88 (2 h). P -values were determined using a t test.

**Techniques Used: **Expressing, Fluorescence, Microscopy, Two Tailed Test, Injection

**Figure Legend Snippet:**(A) Immunofluorescent images of KAP-GFP cells during regeneration in either DMSO or 30 μM BCI. Scale bars are 5 μm. Red arrows point to basal bodies, which are the object of quantification in (D). (B) Western blot for KAP-GFP expression in regenerating cells in either DMSO (D) or 30 μM BCI (B). Total protein was measured with amido black. (C) Quantification of (B). Error bars are the mean with SD for three independent experiments. P -values were calculated using an ordinary one-way ANOVA with Dunnett’s multiple comparisons test. (D) Quantification of KAP-GFP fluorescence at the basal bodies in (A). Error bars are the mean with the 95% confidence interval for the averages from three independent trials (n = 30, N = 3). The P -value was calculated using an unpaired t test with Welch’s correction.

**Techniques Used: **Western Blot, Expressing, Fluorescence

**Figure Legend Snippet:**(A) Western blot of KAP-GFP expression compared with total protein shown with amido black. Cells were treated for 2 h with either 0.5% DMSO, 20 μM BCI, 50 μM MG132, or both 50 μM MG132 and 20 μM BCI. (A, B) Quantification of (A). Error bars are SD of the mean (N = 3). The P -value was determined by an unpaired t test between BCI and BCI + MG132.

**Techniques Used: **Western Blot, Expressing

**Figure Legend Snippet:**BCI inhibits phosphate removal from MAPK. This ultimately alters or inhibits ciliogenesis, KAP-GFP dynamics in cilia, KAP-GFP protein synthesis, NPHP4 protein localization at the transition zone, membrane trafficking, and microtubule organization.

**Techniques Used: **

**Figure Legend Snippet:**Tools and reagents.

**Techniques Used: **Staining, Mutagenesis

# kap gfp (Cell Signaling Technology Inc)

Cell Signaling Technology Inc is a verified supplier

Cell Signaling Technology Inc manufactures this product

## Structured Review

Kap Gfp, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more

https://www.bioz.com/result/kap gfp/product/Cell Signaling Technology Inc

Average 86 stars, based on 1 article reviews

Price from $9.99 to $1999.99

## Images

### 1) Product Images from "ERK pathway activation inhibits ciliogenesis and causes defects in motor behavior, ciliary gating, and cytoskeletal rearrangement"

**Article Title: **ERK pathway activation inhibits ciliogenesis and causes defects in motor behavior, ciliary gating, and cytoskeletal rearrangement

**Journal: **bioRxiv

**doi: **10.1101/2022.05.18.492507

**Figure Legend Snippet:**( A ) Chlamydomonas expressing KAP-GFP. Indicated are the measured areas of KAP-GFP localization: the 2 basal bodies and cilia. Scale bar is 5 µ m. ( B ) Super plot quantification of basal body fluorescence in (A) following a 2 hour treatment with BCI at the indicated concentrations. Error bars are mean with 95% confidence interval. The average from each trial is plotted in larger circles on top of the individual points. P=0.5126 which was determined by an ordinary one-way ANOVA. ( C ) Ciliary length plotted against ciliary fluorescence for cells in DMSO (orange circles) or 30 µ M BCI (purple). ( D ) Example kymographs collected from total internal reflection fluorescent (TIRF) microscopy of KAP-GFP movement in cilia in cells treated with DMSO (top) or 30 µ M BCI (bottom) for 2 hours. Vertical scale bars are 4 µ m. Horizontal scale bars are 2 seconds. ( E-F ) KAP-GFP dynamics quantified from the kymographs (n=20, N=1). The error bars are mean with 95% confidence interval (n=20, N=1). P values were calculated from a two-tailed unpaired t-test for pairwise comparisons. ( E ) Frequency of KAP-GFP trains measured as the total amount of trains counted over the total amount of time the kymograph was collected. For DMSO, n=40 (1 hr and 2 hr). For BCI, n=30 (1 hr) and 34 (2 hr). ( F ) Velocity of KAP-GFP trains measured as the distance traveled in µ m over time in seconds. For DMSO, n=100 (1 hr and 2 hr). For BCI, n=93 (1 hr) and 88 (2 hr). ( G ) Relative injection size KAP-GFP trains measured as relative total fluorescent intensity of each train relative to measurements taken before treatment with DMSO or BCI (data not shown). For DMSO, n=100 (1 hr and 2 hr). For BCI, n=93 (1 hr) and 88 (2 hr).

**Techniques Used: **Expressing, Fluorescence, Microscopy, Two Tailed Test, Injection

**Figure Legend Snippet:**(A) Experimental illustration for (B-E). Chlamydomonas cells were treated for 2 hours with either 0.5% DMSO, 30 µ M BCI to induce partial ciliary resorption, or 45 µ M BCI to induce complete ciliary resorption. (B) Western blot for KAP-GFP expression in the presence of DMSO or 30 µ M BCI for 2 hours. Cilia were isolated and KAP-GFP expression checked separately from the cell bodies using a GFP antibody. Total protein was determined with amido black. ( C ) Intensity quantification of GFP normalized to total protein. Error bars are standard deviation of the mean. P values were determined by a Welch’s t test (N=3). D is DMSO (0.5%), B is BCI (30 µ M). (D) Western blot for KAP-GFP expression in the presence of DMSO or 45 µ M BCI to induce complete ciliary resorption. GFP antibody was used to probe for KAP-GFP. Total protein was determined with Coomassie Brilliant Blue. ( E ) Intensity quantification (right) measures protein from 3 independent experiments. Error bars are mean with standard deviation. (F) Immunofluorescent images of KAP-GFP cells during regeneration in either 0.5% DMSO or 30 µ M BCI. Scale bars are 5 µ m. Red arrows point to basal bodies which are the object of quantification in (G). (G) Quantification of KAP-GFP fluorescence at the basal bodies. Error bars are mean with 95% confidence interval of the averages for 3 independent trials (n=30, N=3). The P value was calculated using an unpaired t-test with Welch’s correction. (H) Western blot for KAP-GFP expression in regenerating cells in either DMSO or 30 µ M BCI. Total protein was measured with amido black. (I) Quantification of F. Error bars are mean with standard deviation for 3 independent experiments. P values were calculated using an ordinary one-way ANOVA with Dunnett’s multiple comparisons test. (J) Double deciliation with a 1 hour regrowth in cycloheximide (CHX). After the first pH shock, cycloheximide (CHX) is added for the first 60 minutes. Cells were deciliated once more, regenerated in 0.2% DMSO or 20 µ M BCI for 4 hours, and washed out or not and grown for 4 more hours. Error bars are 95% confidence interval on the means of 3 experiments (n=30, N=3). P values were determined with a two-way ANOVA with Tukey’s correction for multiple comparisons.

**Techniques Used: **Western Blot, Expressing, Isolation, Standard Deviation, Fluorescence

**Figure Legend Snippet:**(A ) Western blot of KAP-GFP expression compared to total protein exposed with amido black. Cells were treated for 2 hours with either 0.5% DMSO, 30 µ M BCI, 50 µ M MG132, or both 50 µ M MG132 and 30 µ MBCI. ( B ) Quantification of (A). Error bars are standard deviation of the mean (n=1, N=3). The P value was determined by an unpaired t-test between BCI and BCI+MG132. (C) Single regeneration performed in parallel with . Error bars are mean with 95% confidence interval (n=30, N=3). P values were determined with a two-way ANOVA with Tukey’s correction for multiple comparisons.

**Techniques Used: **Western Blot, Expressing, Standard Deviation